Quinoxaline derivatives useful as FGFR kinase modulators

ABSTRACT

The invention relates to new quinoxaline derivative compounds, to pharmaceutical compositions comprising said compounds, to processes for the preparation of said compounds and to the use of said compounds in the treatment of diseases, e.g. cancer.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.15/128,345, filed on Sep. 22, 2016 (published as US 2017-0101396 A1 onApr. 13, 2017), which is a national stage filing under section 371 ofInternational Application No. PCT/EP2015/056507, filed on Mar. 26, 2015,and published as WO 2015/144803 on Oct. 1, 2015, and claims priority toEuropean Patent Application No. 14161820.7 filed on Mar. 26, 2014. Theentire disclosures of each of the prior applications are herebyincorporated herein by reference.

FIELD OF THE INVENTION

The invention relates to new quinoxaline derivative compounds, topharmaceutical compositions comprising said compounds, to processes forthe preparation of said compounds and to the use of said compounds inthe treatment of diseases, e.g. cancer.

SUMMARY OF THE INVENTION

According to a first aspect of the invention there is provided compoundsof formula (I):

including any tautomeric or stereochemically isomeric form thereof,whereinn represents an integer equal to 1 or 2;R₁ represents hydrogen, C₁₋₆alkyl, hydroxyC₁₋₆alkyl, C₁₋₆alkylsubstituted with —C(═O)NHCH₃, or C₁₋₆alkyl substituted with—S(═O)₂—C₁₋₄alkyl;R_(2a) represents hydrogen, fluoro or chloro;R_(2b) or R_(2c) each independently represent methoxy or hydroxyl;R₃ represent hydrogen, C₁₋₆alkyl, C₃₋₆cycloalkyl, or C₁₋₂alkylsubstituted with C₃₋₆cycloalkyl;R₄ represents hydrogen, methyl or ethyl;the pharmaceutically acceptable salts thereof or the solvates thereof.

In one embodiment there is provided compounds of formula (Ia):

including any tautomeric or stereochemically isomeric form thereof,wherein n represents an integer equal to 1 or 2;R₁ represents hydrogen, C₁₋₆ alkyl, hydroxyC₁₋₆ alkyl, C₁₋₆ alkylsubstituted with —C(═O)NHCH₃, or C₁₋₆alkyl substituted with—S(═O)₂—C₁₋₄alkyl;R_(2a) represents hydrogen, fluoro or chloro;R_(2b) or R_(2c) each independently represent methoxy or hydroxyl;R₃ represent hydrogen, C₁₋₆alkyl, C₃₋₆cycloalkyl, or C₁₋₂alkylsubstituted with C₃₋₆cycloalkyl; the pharmaceutically acceptable saltsthereof or the solvates thereof.

In one embodiment there is provided compounds of formula (Ib):

including any tautomeric or stereochemically isomeric form thereof,whereinn represents an integer equal to 1 or 2;R₁ represents hydrogen, C₁₋₆alkyl, hydroxyC₁₋₆alkyl, C₁₋₆alkylsubstituted with —C(═O)NHCH₃, or C₁₋₆alkyl substituted with—S(═O)₂—C₁₋₄alkyl;R_(2b) or R_(2c) each independently represent methoxy or hydroxyl;R₃ represent hydrogen, C₁₋₆alkyl, C₃₋₆cycloalkyl, or C₁₋₂alkylsubstituted with C₃₋₆cycloalkyl; the pharmaceutically acceptable saltsthereof or the solvates thereof.

In one embodiment there is provided compounds of formula (Ic):

including any tautomeric or stereochemically isomeric form thereof,whereinR₁ represents hydrogen, C₁₋₆ alkyl, hydroxyC₁₋₆ alkyl, C₁₋₆ alkylsubstituted with —C(═O)NHCH₃, or C₁₋₆alkyl substituted with—S(═O)₂—C₁₋₄alkyl;R_(2b) or R_(2c) each independently represent methoxy or hydroxyl;the pharmaceutically acceptable salts thereof or the solvates thereof.

In one embodiment there is provided compounds of formula (Id):

including any tautomeric or stereochemically isomeric form thereof,whereinR_(2b) or R₂, each independently represent methoxy or hydroxyl;the pharmaceutically acceptable salts thereof or the solvates thereof.

In one embodiment there is provided compounds of formula (Ie):

including any tautomeric or stereochemically isomeric form thereof,whereinR₁ represents hydrogen, C₁₋₆ alkyl, hydroxyC₁₋₆ alkyl, C₁₋₆ alkylsubstituted with —C(═O)NHCH₃, or C₁₋₆alkyl substituted with—S(═O)₂—C₁₋₄alkyl;R_(2b) or R₂, each independently represent methoxy or hydroxyl;the pharmaceutically acceptable salts thereof or the solvates thereof

WO2006/092430, WO2008/003702, WO01/68047, WO2005/007099, WO2004/098494,WO2009/141386, WO2004/030635, WO2008/141065, WO2011/026579,WO2011/028947, WO2007/003419, WO00/42026, WO2012/154760, WO2011/047129,WO2003/076416, WO2002/096873, WO2000/055153, EP548934, U.S. Pat. No.4,166,117, WO2011/135376, WO2012/073017, WO2013/061074, WO2013/061081,WO2013/061077, WO2013/061080, WO2013/179034, WO2013/179033,WO2014/174307 which each disclose a series of heterocyclyl derivatives.

DETAILED DESCRIPTION OF THE INVENTION

Unless the context indicates otherwise, references to formula (I) in allsections of this document (including the uses, methods and other aspectsof the invention) include references to all other sub-formula (e.g. Ia,Ib, Ic, Id), sub-groups, preferences, embodiments and examples asdefined herein.

The prefix “C_(x-y)” (where x and y are integers) as used herein refersto the number of carbon atoms in a given group. Thus, a C₁₋₆alkyl groupcontains from 1 to 6 carbon atoms, a C₃₋₆cycloalkyl group contains from3 to 6 carbon atoms, a hydroxyC₁₋₆ alkyl group contains from 1 to 6carbon atoms, and so on.

The term ‘C₁₋₂alkyl’, ‘C₁₋₄alky’, or ‘C₁₋₆alkyl’ as used herein as agroup or part of a group refers to a linear or branched saturatedhydrocarbon group containing 1 or 2, or from 1 to 4 or 1 to 6 carbonatoms. Examples of such groups include methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl,isopentyl, neopentyl or hexyl and the like.

The term ‘C₃₋₆cycloalkyl’ as used herein refers to a saturatedmonocyclic hydrocarbon ring of 3 to 6 carbon atoms. Examples of suchgroups include cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl.

The term ‘hydroxyC₁₋₄alkyl’ or ‘hydroxyC₁₋₆alky’ as used herein as agroup or part of a group refers to a C₁₋₄ alkyl or C₁₋₆alkyl group asdefined herein wherein one or more than one hydrogen atom is replacedwith a hydroxyl group. The terms ‘hydroxyC₁₋₄alkyl’ or‘hydroxyC₁₋₆alkyl’ therefore include monohydroxyC₁₋₄alkykmonohydroxyC₁₋₆alkyl and also polyhydroxyC₁₋₄alkyl andpolyhydroxyC₁₋₆alkyl. There may be one, two, three or more hydrogenatoms replaced with a hydroxyl group, so the hydroxyC₁₋₄alkyl orhydroxyC₁₋₆alkyl may have one, two, three or more hydroxyl groups.Examples of such groups include hydroxymethyl, hydroxyethyl,hydroxypropyl and the like.

In one embodiment, in a compound of formula (I), n represents an integerequal to 1.

In one embodiment, in a compound of formula (I), n represents an integerequal to 2.

In one embodiment, in a compound of formula (I), R₁ represents hydrogenor C₁₋₆ alkyl, in particular C₁₋₆alkyl, more in particular methyl.

In one embodiment, in a compound of formula (I), R₁ represents hydrogenor C₁₋₆ alkyl, in particular C₁₋₆alkyl, more in particular ethyl.

In one embodiment, in a compound of formula (I), R₁ represents hydrogen.

In one embodiment, in a compound of formula (I), R_(2a) representshydrogen or fluoro.

In one embodiment, in a compound of formula (I), R_(2a) representshydrogen.

In one embodiment, in a compound of formula (I), R_(2a) representsfluoro.

In one embodiment, in a compound of formula (I), R_(2b) representsmethoxy.

In one embodiment, in a compound of formula (I), R_(2b) representshydroxy.

In one embodiment, in a compound of formula (I), R_(2c) representsmethoxy.

In one embodiment, in a compound of formula (I), R_(2c) representshydroxy.

In one embodiment, in a compound of formula (I), R_(2b) representsmethoxy and R_(2c) represents hydroxyl.

In one embodiment, in a compound of formula (I), R_(2b) representshydroxyl and R_(2c) represents methoxy.

In one embodiment, in a compound of formula (I), R_(2b) and R_(2c) bothrepresent methoxy.

In one embodiment, in a compound of formula (I), R_(2b) and R_(2c) bothrepresent hydroxyl.

In one embodiment, in a compound of formula (I), R₃ represents hydrogen.

In one embodiment, in a compound of formula (I), R₃ represents C₁₋₆alkyl, in particular C₁₋₄ alkyl, even more in particular isopropyl.

In one embodiment, in a compound of formula (I), R₃ represents C₁₋₆alkyl, in particular C₁₋₄ alkyl, even more in particular methyl.

In one embodiment, in a compound of formula (I), R₄ represents hydrogen.

In one embodiment, in a compound of formula (I), R₄ represents methyl orethyl.

In one embodiment, in a compound of formula (I),

n represents an integer equal to 1;

R₁ represents C₁₋₆alkyl, in particular C₁₋₄ alkyl, more in particularmethyl;

R_(2a) represents hydrogen or fluoro, in particular hydrogen;

R_(2b) represents methoxy;

R_(2c) represents methoxy;

R₃ represents hydrogen or C₁₋₆ alkyl, in particular C₁₋₆ alkyl, more inparticular C₁₋₄ alkyl, even more in particular isopropyl;

R₄ represents hydrogen.

In one embodiment, in a compound of formula (I),

n represents an integer equal to 1 or 2;

R₁ represents C₁₋₆ alkyl, in particular C₁₋₄ alkyl, more in particularmethyl or ethyl;

R_(2a) represents hydrogen or fluoro, in particular hydrogen;

R_(2b) represents methoxy;

R_(2c) represents methoxy;

R₃ represents hydrogen or C₁₋₆ alkyl, in particular C₁₋₆ alkyl, more inparticular C₁₋₄ alkyl, even more in particular isopropyl or methyl;

R₄ represents hydrogen.

In one embodiment, in a compound of formula (Ia), n represents aninteger equal to 1.

In one embodiment, in a compound of formula (Ia), n represents aninteger equal to 2.

In one embodiment, in a compound of formula (Ia), R₁ represents hydrogenor C₁₋₆alkyl, in particular C₁₋₆alkyl, more in particular methyl.

In one embodiment, in a compound of formula (Ia), R₁ represents hydrogenor C₁₋₆ alkyl, in particular C₁₋₆alkyl, more in particular ethyl.

In one embodiment, in a compound of formula (Ia), R₁ representshydrogen.

In one embodiment, in a compound of formula (Ia), R_(2a) representshydrogen or fluoro.

In one embodiment, in a compound of formula (Ia), R_(2a) representshydrogen.

In one embodiment, in a compound of formula (Ia), R_(2a) representsfluoro.

In one embodiment, in a compound of formula (Ia), R_(2b) representsmethoxy.

In one embodiment, in a compound of formula (Ia), R_(2b) representshydroxy.

In one embodiment, in a compound of formula (Ia), R_(2c) representsmethoxy.

In one embodiment, in a compound of formula (Ia), R_(2c) representshydroxy.

In one embodiment, in a compound of formula (Ia), R_(2b) representsmethoxy and R_(2c) represents hydroxyl.

In one embodiment, in a compound of formula (Ia), R_(2b) representshydroxyl and R_(2c) represents methoxy.

In one embodiment, in a compound of formula (Ia), R_(2b) and R_(2c) bothrepresent methoxy.

In one embodiment, in a compound of formula (Ia), R_(2b) and R_(2c) bothrepresent hydroxyl.

In one embodiment, in a compound of formula (Ia), R₃ representshydrogen.

In one embodiment, in a compound of formula (Ia), R₃ represents C₁₋₆alkyl, in particular C₁₋₄alkyl, even more in particular isopropyl.

In one embodiment, in a compound of formula (Ia), R₃ represents C₁₋₆alkyl, in particular C₁₋₄alkyl, even more in particular methyl.

In one embodiment, in a compound of formula (Ia),

n represents an integer equal to 1;

R₁ represents C₁₋₆alkyl, in particular C₁₋₄alkyl, more in particularmethyl;

R_(2a) represents hydrogen or fluoro, in particular hydrogen;

R_(2b) represents methoxy;

R_(2c) represents methoxy;

R₃ represents hydrogen or C₁₋₆ alkyl, in particular C₁₋₆ alkyl, more inparticular C₁₋₄ alkyl, even more in particular isopropyl.

In one embodiment, in a compound of formula (Ia),

n represents an integer equal to 1 or 2;

R₁ represents C₁₋₆alkyl, in particular C₁₋₄alkyl, more in particularmethyl or ethyl;

R_(2a) represents hydrogen or fluoro, in particular hydrogen;

R_(2b) represents methoxy;

R_(2c) represents methoxy;

R₃ represents hydrogen or C₁₋₆ alkyl, in particular C₁₋₆ alkyl, more inparticular C₁₋₄ alkyl, even more in particular isopropyl or methyl.

In one embodiment, in a compound of formula (Ib), n represents aninteger equal to 1.

In one embodiment, in a compound of formula (Ib), n represents aninteger equal to 2.

In one embodiment, in a compound of formula (Ib), R₁ represents hydrogenor C₁₋₆ alkyl, in particular C₁₋₆alkyl, more in particular methyl.

In one embodiment, in a compound of formula (Ib), R₁ represents hydrogenor C₁₋₆alkyl, in particular C₁₋₆alkyl, more in particular ethyl.

In one embodiment, in a compound of formula (Ib), R₁ representshydrogen.

In one embodiment, in a compound of formula (Ib), R_(2b) representsmethoxy.

In one embodiment, in a compound of formula (Ib), R_(2b) representshydroxy.

In one embodiment, in a compound of formula (Ib), R_(2c) representsmethoxy.

In one embodiment, in a compound of formula (Ib), R_(2c) representshydroxy.

In one embodiment, in a compound of formula (Ib), R_(2b) representsmethoxy and R_(2c) represents hydroxyl.

In one embodiment, in a compound of formula (Ib), R_(2b) representshydroxyl and R_(2c) represents methoxy.

In one embodiment, in a compound of formula (Ib), R_(2b) and R_(2c) bothrepresent methoxy.

In one embodiment, in a compound of formula (Ib), R_(2b) and R_(2c) bothrepresent hydroxyl.

In one embodiment, in a compound of formula (Ib), R₃ representshydrogen.

In one embodiment, in a compound of formula (Ib), R₃ represents C₁₋₆alkyl, in particular C₁₋₄alkyl, even more in particular isopropyl.

In one embodiment, in a compound of formula (Ib), R₃ represents C₁₋₆alkyl, in particular C₁₋₄alkyl, even more in particular methyl.

In one embodiment, in a compound of formula (Ib),

n represents an integer equal to 1;

R₁ represents C₁₋₆ alkyl, in particular C₁₋₄alkyl, more in particularmethyl;

R_(2b) represents methoxy;

R_(2c) represents methoxy;

R₃ represents hydrogen or C₁₋₆ alkyl, in particular C₁₋₆ alkyl, more inparticular C₁₋₄ alkyl, even more in particular isopropyl.

In one embodiment, in a compound of formula (Ib),

n represents an integer equal to 1 or 2;

R₁ represents C₁₋₆ alkyl, in particular C₁₋₄alkyl, more in particularmethyl or ethyl;

R_(2b) represents methoxy;

R_(2c) represents methoxy;

R₃ represents hydrogen or C₁₋₆ alkyl, in particular C₁₋₆ alkyl, more inparticular C₁₋₄ alkyl, even more in particular isopropyl or methyl.

In one embodiment, in a compound of formula (Ic), R₁ represents hydrogenor C₁₋₆ alkyl, in particular C₁₋₆alkyl, more in particular methyl.

In one embodiment, in a compound of formula (Ic), R₁ represents hydrogenor C₁₋₆ alkyl, in particular C₁₋₆alkyl, more in particular ethyl.

In one embodiment, in a compound of formula (Ic), R₁ representshydrogen.

In one embodiment, in a compound of formula (Ic), R_(2b) representsmethoxy.

In one embodiment, in a compound of formula (Ic), R_(2b) representshydroxy.

In one embodiment, in a compound of formula (Ic), R_(2c) representsmethoxy.

In one embodiment, in a compound of formula (Ic), R_(2c) representshydroxy.

In one embodiment, in a compound of formula (Ic), R_(2b) representsmethoxy and R_(2c) represents hydroxyl.

In one embodiment, in a compound of formula (Ic), R_(2b) representshydroxyl and R_(2c) represents methoxy.

In one embodiment, in a compound of formula (Ic), R_(2b) and R_(2c) bothrepresent methoxy.

In one embodiment, in a compound of formula (Ic), R_(2b) and R_(2c) bothrepresent hydroxyl.

In one embodiment, in a compound of formula (Ic),

R₁ represents C₁₋₆alkyl, in particular C₁₋₄alkyl, more in particularmethyl;

R_(2b) represents methoxy;

R_(2c) represents methoxy.

In one embodiment, in a compound of formula (Ic),

R₁ represents C₁₋₆alkyl, in particular C₁₋₄alkyl, more in particularmethyl or ethyl;

R_(2b) represents methoxy;

R_(2c) represents methoxy.

In one embodiment, in a compound of formula (Id), R_(2b) representsmethoxy.

In one embodiment, in a compound of formula (Id), R_(2b) representshydroxy.

In one embodiment, in a compound of formula (Id), R_(2c) representsmethoxy.

In one embodiment, in a compound of formula (Id), R_(2c) representshydroxy.

In one embodiment, in a compound of formula (Id), R_(2b) representsmethoxy and R_(2c) represents hydroxyl.

In one embodiment, in a compound of formula (Id), R_(2b) representshydroxyl and R_(2c) represents methoxy.

In one embodiment, in a compound of formula (Id), R_(2b) and R_(2c) bothrepresent methoxy.

In one embodiment, in a compound of formula (Id), R_(2b) and R_(2c) bothrepresent hydroxyl.

In one embodiment, in a compound of formula (Ie), R₁ represents hydrogenor C₁₋₆ alkyl, in particular C₁₋₆alkyl, more in particular methyl.

In one embodiment, in a compound of formula (Ie), R₁ represents hydrogenor C₁₋₆ alkyl, in particular C₁₋₆alkyl, more in particular ethyl.

In one embodiment, in a compound of formula (Ie), R₁ representshydrogen.

In one embodiment, in a compound of formula (Ie), R_(2b) representsmethoxy.

In one embodiment, in a compound of formula (Ie), R_(2b) representshydroxy.

In one embodiment, in a compound of formula (Ie), R_(2c) representsmethoxy.

In one embodiment, in a compound of formula (Ie), R_(2c) representshydroxy.

In one embodiment, in a compound of formula (Ie), R_(2b) representsmethoxy and R_(2c) represents hydroxyl.

In one embodiment, in a compound of formula (Ie), R_(2b) representshydroxyl and R_(2c) represents methoxy.

In one embodiment, in a compound of formula (Ie), R_(2b) and R_(2c) bothrepresent methoxy.

In one embodiment, in a compound of formula (Ie), R_(2b) and R_(2c) bothrepresent hydroxyl.

In one embodiment, in a compound of formula (Ie),

R₁ represents C₁₋₆ alkyl, in particular C₁₋₄alkyl, more in particularmethyl;

R_(2b) represents methoxy;

R_(2c) represents methoxy.

In one embodiment, in a compound of formula (Ie),

R₁ represents C₁₋₆ alkyl, in particular C₁₋₄alkyl, more in particularmethyl or ethyl;

R_(2b) represents methoxy;

R_(2c) represents methoxy.

In one embodiment, in a compound of formula (I),

n represents an integer equal to 1 or 2;

R₁ represents hydrogen or C₁₋₆ alkyl, in particular C₁₋₄alkyl, more inparticular methyl or ethyl;

R_(2a) represents hydrogen or fluoro, in particular hydrogen;

R_(2b) represents methoxy or hydroxyl;

R_(2c) represents methoxy or hydroxyl;

R₃ represents C₁₋₆ alkyl, more in particular C₁₋₄ alkyl, even more inparticular isopropyl or methyl;

R₄ represents hydrogen.

In one embodiment, the compound of formula (I) as defined herein isselected from the following compounds or is one of the followingcompounds:

a pharmaceutically acceptable salt thereof or a solvate thereof.

In one embodiment, the compound of formula (I) as defined herein isselected from the following compounds or is one of the followingcompounds:

a pharmaceutically acceptable salt thereof or a solvate thereof.

For the avoidance of doubt, it is to be understood that each general andspecific preference, embodiment and example for one substituent may becombined with each general and specific preference, embodiment andexample for one or more, preferably, all other substituents as definedherein and that all such embodiments are embraced by this application.

Methods for the Preparation of Compounds of Formula (I)

In this section, as in all other sections of this application unless thecontext indicates otherwise, references to formula (I) also include allother sub-groups and examples thereof as defined herein.

In general, compounds of formula (I) can be prepared according to thefollowing reaction scheme

In Scheme 1, the following reaction conditions apply:

1: reaction of an intermediate of formula (II) with formaldehyde in thepresence of a suitable solvent, such as for example dioxane,N,N-dimethylformamide, N,N-dimethylacetamide, at a temperature rangingfrom room temperature to reflux.

It is considered to be within the knowledge of the person skilled in theart to recognize in which condition and on which part of the molecule aprotective group may be appropriate. For instance, protective group onthe R₁ substituent or on the pyrrazole moiety, or protective group onthe R₃ substituent or on the R_(2a,b,c) substituent or combinationsthereof. The skilled person is also considered to be able to recognizethe most feasible protective group, such as for example—C(═O)—O—C₁₋₄alkyl or

or O—Si(CH₃)₂(C(CH₃)₃) or —CH₂—O—CH₂CH₂—O—CH₃.

The present invention also comprises deuterated compounds. Thesedeuterated compounds may be prepared by using the appropriate deuteratedintermediates during the synthesis process.

The compounds of formula (I) may also be converted into each other viaart-known reactions or functional group transformations.

Compounds of formula (I) wherein R₁ represents hydrogen can be convertedinto a compound of formula (I) wherein R₁ represents C₁₋₆alkyl orhydroxyC₁₋₆alkyl, by reaction with C₁₋₆alkyl-W or hydroxyC₁₋₆ alkyl-W,wherein W represents a suitable leaving group, such as for example halo,e.g. bromo and the like, in the presence of a suitable base, such as forexample sodium hydride or potassium carbonate, and a suitable solvent,such as for example acetonitrile or N,N-dimethylformamide.

Compounds of formula (I) wherein R₁ represents hydrogen can also beconverted into a compound of formula (I) wherein R₁ represents C₁₋₆alkyl-OH, by reaction with W—C₁₋₆alkyl-O—Si(CH₃)₂(C(CH₃)₃) in thepresence of a suitable base, such as for example sodium hydride, and asuitable solvent, such as for example N,N-dimethylformamide, followed bya deprotection reaction of the silyl protecting group by art-knownmethods.

Compounds of formula (I) wherein R₁ represents hydrogen, can also beconverted into a compound of formula (I) wherein R₁ represents ethylsubstituted with —S(═O)₂—C₁₋₆alkyl, by reaction withC₁₋₆alkyl-vinylsulfone, in the presence of a suitable base, such as forexample triethylamine, and a suitable solvent, such as for example analcohol, e.g. methanol or by reaction with C₁₋₆alkyl-2-bromoethylsulfonein the presence of a suitable deprotonating agent, such as for exampleNaH, and a suitable solvent, such as for example dimethylformamide.Compounds of formula (I) wherein R_(2b) or R_(2c) represents —OCH₃ canbe converted into a compound of formula (I) wherein R_(2b) or R_(2c)represents —OH by reaction with boron tribromide in the presence of asuitable solvent, such as for example dichloromethane.

Compounds of formula (I) wherein R_(2b) or R_(2c) represents —OH can beconverted into a compound of formula (I) wherein R_(2b) or R_(2c)represents —OCH₃ by reaction with methyl iodine in the presence of asuitable base, such as for example potassium carbonate, and a suitablesolvent, such as for example N,N-dimethylformamide.

In general, compounds of formula (Id) can also be prepared by incubatingthem with liver fractions of animals, e.g. rat or human, andsubsequently isolating the desired products from the incubation mediumaccording to the following reaction scheme.

Intermediates of formula (II) or (II-a) can be prepared as described inWO2011/135376 (for instance compounds of formula (I-b) or (I-b-3) ofWO2011/135376).

A further aspect of the invention is a process for the preparation of acompound of formula (I) as defined herein, which process comprises:

(i) reacting a compound of formula (II) with formaldehyde in thepresence of a suitable solvent, such as for example dioxane,N,N-dimethylformamide, N,N-dimethylacetamide, at a suitable temperature,such as a temperature ranging from room temperature to reflux;

-   -   wherein R₁, R_(2a), R_(2b), R_(2c), R₃, R₄ and n are as defined        herein; and optionally thereafter converting one compound of the        formula (I) into another compound of the formula (I).        Pharmaceutically Acceptable Salts, Solvates or Derivatives        Thereof

In this section, as in all other sections of this application, unlessthe context indicates otherwise, references to formula (I) includereferences to all other sub-groups, preferences, embodiments andexamples thereof as defined herein.

Unless otherwise specified, a reference to a particular compound alsoincludes ionic forms, salts, solvates, isomers, tautomers, esters,prodrugs, isotopes and protected forms thereof, for example, asdiscussed below; preferably, the ionic forms, or salts or tautomers orisomers or solvates thereof; and more preferably, the ionic forms, orsalts or tautomers or solvates or protected forms thereof, even morepreferably the salts or tautomers or solvates thereof. Many compounds ofthe formula (I) can exist in the form of salts, for example acidaddition salts or, in certain cases salts of organic and inorganic basessuch as carboxylate, sulphonate and phosphate salts. All such salts arewithin the scope of this invention, and references to compounds of theformula (I) include the salt forms of the compounds. It will beappreciated that references to “derivatives” include references to ionicforms, salts, solvates, isomers, tautomers, esters, prodrugs, isotopesand protected forms thereof.

According to one aspect of the invention there is provided a compound asdefined herein or a salt, tautomer, or solvate thereof. According to afurther aspect of the invention there is provided a compound as definedherein or a salt or solvate thereof. References to compounds of theformula (I) and sub-groups thereof as defined herein include withintheir scope the salts or solvates or tautomers of the compounds.

The salt forms of the compounds of the invention are typicallypharmaceutically acceptable salts, and examples of pharmaceuticallyacceptable salts are discussed in Berge et al. (1977) “PharmaceuticallyAcceptable Salts,” J. Pharm. Sci., Vol. 66, pp. 1-19. However, saltsthat are not pharmaceutically acceptable may also be prepared asintermediate forms which may then be converted into pharmaceuticallyacceptable salts. Such non-pharmaceutically acceptable salts forms,which may be useful, for example, in the purification or separation ofthe compounds of the invention, also form part of the invention.

The salts of the present invention can be synthesized from the parentcompound that contains a basic or acidic moiety by conventional chemicalmethods such as methods described in Pharmaceutical Salts: Properties,Selection, and Use, P. Heinrich Stahl (Editor), Camille G. Wermuth(Editor), ISBN: 3-90639-026-8, Hardcover, 388 pages, August 2002.Generally, such salts can be prepared by reacting the free acid or baseforms of these compounds with the appropriate base or acid in water orin an organic solvent, or in a mixture of the two; generally, nonaqueousmedia such as ether, ethyl acetate, ethanol, isopropanol, oracetonitrile are used. The compounds of the invention may exist as mono-or di-salts depending upon the pKa of the acid from which the salt isformed.

Acid addition salts may be formed with a wide variety of acids, bothinorganic and organic. Examples of acid addition salts include saltsformed with an acid selected from the group consisting of acetic,2,2-dichloroacetic, adipic, alginic, ascorbic (e.g. L-ascorbic),L-aspartic, benzenesulphonic, benzoic, 4-acetamidobenzoic, butanoic, (+)camphoric, camphor-sulphonic, (+)-(1S)-camphor-10-sulphonic, capric,caproic, caprylic, cinnamic, citric, cyclamic, dodecyl sulphuric,ethane-1,2-di sulphonic, ethanesulphonic, 2-hydroxyethanesulphonic,formic, fumaric, galactaric, gentisic, glucoheptonic, D-gluconic,glucuronic (e.g. D-glucuronic), glutamic (e.g. L-glutamic),α-oxoglutaric, glycolic, hippuric, hydrobromic, hydrochloric, hydriodic,isethionic, lactic (e.g. (+)-L-lactic, (±)-DL-lactic), lactobionic,maleic, malic, (−)-L-malic, malonic, (±)-DL-mandelic, methanesulphonic,naphthalenesulphonic (e.g. naphthalene-2-sulphonic),naphthalene-1,5-disulphonic, 1-hydroxy-2-naphthoic, nicotinic, nitric,oleic, orotic, oxalic, palmitic, pamoic, phosphoric, propionic,L-pyroglutamic, pyruvic, salicylic, 4-amino-salicylic, sebacic, stearic,succinic, sulphuric, tannic, (+)-L-tartaric, thiocyanic,toluenesulphonic (e.g. p-toluenesulphonic), undecylenic and valericacids, as well as acylated amino acids and cation exchange resins.

One particular group of salts consists of salts formed from acetic,hydrochloric, hydriodic, phosphoric, nitric, sulphuric, citric, lactic,succinic, maleic, malic, isethionic, fumaric, benzenesulphonic,toluenesulphonic, methanesulphonic (mesylate), ethanesulphonic,naphthalenesulphonic, valeric, acetic, propanoic, butanoic, malonic,glucuronic and lactobionic acids. Another group of acid addition saltsincludes salts formed from acetic, adipic, ascorbic, aspartic, citric,DL-Lactic, fumaric, gluconic, glucuronic, hippuric, hydrochloric,glutamic, DL-malic, methanesulphonic, sebacic, stearic, succinic andtartaric acids.

If the compound is anionic, or has a functional group which may beanionic, then a salt may be formed with a suitable cation. Examples ofsuitable inorganic cations include, but are not limited to, alkali metalions such as Na⁺ and K⁺, alkaline earth metal cations such as Ca²⁺ andMg²⁺, and other cations such as Al³⁺. Examples of suitable organiccations include, but are not limited to, ammonium ion (i.e., NH₄ ⁺) andsubstituted ammonium ions (e.g., NH₃R⁺, NH₂R₂ ⁺, NHR₃ ⁺, NR₄ ⁺).

Examples of some suitable substituted ammonium ions are those derivedfrom: ethylamine, diethylamine, dicyclohexylamine, triethylamine,butylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine,benzylamine, phenylbenzylamine, choline, meglumine, and tromethamine, aswell as amino acids, such as lysine and arginine. An example of a commonquaternary ammonium ion is N(CH₃)₄ ⁺.

Where the compounds of the formula (I) contain an amine function, thesemay form quaternary ammonium salts, for example by reaction with analkylating agent according to methods well known to the skilled person.Such quaternary ammonium compounds are within the scope of formula (I).Compounds of the formula (I) containing an amine function may also formN-oxides. A reference herein to a compound of the formula (I) thatcontains an amine function also includes the N-oxide. Where a compoundcontains several amine functions, one or more than one nitrogen atom maybe oxidised to form an N-oxide. Particular examples of N-oxides are theN-oxides of a tertiary amine or a nitrogen atom of a nitrogen-containingheterocycle. N-Oxides can be formed by treatment of the correspondingamine with an oxidizing agent such as hydrogen peroxide or a per-acid(e.g. a peroxycarboxylic acid), see for example Advanced OrganicChemistry, by Jerry March, 4^(th) Edition, Wiley Interscience, pages.More particularly, N-oxides can be made by the procedure of L. W. Deady(Syn. Comm. (1977), 7, 509-514) in which the amine compound is reactedwith m-chloroperoxybenzoic acid (MCPBA), for example, in an inertsolvent such as dichloromethane.

The compounds of the invention may form solvates, for example with water(i.e., hydrates) or common organic solvents. As used herein, the term“solvate” means a physical association of the compounds of the presentinvention with one or more solvent molecules. This physical associationinvolves varying degrees of ionic and covalent bonding, includinghydrogen bonding. In certain instances the solvate will be capable ofisolation, for example when one or more solvent molecules areincorporated in the crystal lattice of the crystalline solid. The term“solvate” is intended to encompass both solution-phase and isolatablesolvates. Non-limiting examples of suitable solvates include compoundsof the invention in combination with water, isopropanol, ethanol,methanol, DMSO, ethyl acetate, acetic acid or ethanolamine and the like.The compounds of the invention may exert their biological effects whilstthey are in solution.

Solvates are well known in pharmaceutical chemistry. They can beimportant to the processes for the preparation of a substance (e.g. inrelation to their purification, the storage of the substance (e.g. itsstability) and the ease of handling of the substance and are oftenformed as part of the isolation or purification stages of a chemicalsynthesis. A person skilled in the art can determine by means ofstandard and long used techniques whether a hydrate or other solvate hasformed by the isolation conditions or purification conditions used toprepare a given compound. Examples of such techniques includethermogravimetric analysis (TGA), differential scanning calorimetry(DSC), X-ray crystallography (e.g. single crystal X-ray crystallographyor X-ray powder diffraction) and Solid State NMR (SS-NMR, also known asMagic Angle Spinning NMR or MAS-NMR). Such techniques are as much a partof the standard analytical toolkit of the skilled chemist as NMR, IR,HPLC and MS. Alternatively the skilled person can deliberately form asolvate using crystallisation conditions that include an amount of thesolvent required for the particular solvate. Thereafter the standardmethods described above, can be used to establish whether solvates hadformed. Also encompassed by formula (I) are any complexes (e.g.inclusion complexes or clathrates with compounds such as cyclodextrins,or complexes with metals) of the compounds.

Furthermore, the compounds of the present invention may have one or morepolymorph (crystalline) or amorphous forms and as such are intended tobe included in the scope of the invention.

Compounds of the formula (I) may exist in a number of differentgeometric isomeric, and tautomeric forms and references to compounds ofthe formula (I) include all such forms. For the avoidance of doubt,where a compound can exist in one of several geometric isomeric ortautomeric forms and only one is specifically described or shown, allothers are nevertheless embraced by formula (I). Other examples oftautomeric forms include, for example, keto-, enol-, and enolate-forms,as in, for example, the following tautomeric pairs: keto/enol(illustrated below), imine/enamine, amide/imino alcohol,amidine/enediamines, nitroso/oxime, thioketone/enethiol andnitro/aci-nitro.

Where compounds of the formula (I) contain one or more chiral centres,and can exist in the form of two or more optical isomers, references tocompounds of the formula (I) include all optical isomeric forms thereof(e.g. enantiomers, epimers and diastereoisomers), either as individualoptical isomers, or mixtures (e.g. racemic mixtures) of two or moreoptical isomers, unless the context requires otherwise. The opticalisomers may be characterised and identified by their optical activity(i.e. as + and − isomers, or d and 1 isomers) or they may becharacterised in terms of their absolute stereochemistry using the “Rand S” nomenclature developed by Cahn, Ingold and Prelog, see AdvancedOrganic Chemistry by Jerry March, 4^(th) Edition, John Wiley & Sons, NewYork, 1992, pages 109-114, and see also Cahn, Ingold & Prelog (1966)Angew. Chem. Int. Ed. Engl., 5, 385-415. Optical isomers can beseparated by a number of techniques including chiral chromatography(chromatography on a chiral support) and such techniques are well knownto the person skilled in the art. As an alternative to chiralchromatography, optical isomers can be separated by formingdiastereoisomeric salts with chiral acids such as (+)-tartaric acid,(−)-pyroglutamic acid, (−)-di-toluoyl-L-tartaric acid, (+)-mandelicacid, (−)-malic acid, and (−)-camphorsulphonic, separating thediastereoisomers by preferential crystallisation, and then dissociatingthe salts to give the individual enantiomer of the free base.

Where compounds of the formula (I) exist as two or more optical isomericforms, one enantiomer in a pair of enantiomers may exhibit advantagesover the other enantiomer, for example, in terms of biological activity.Thus, in certain circumstances, it may be desirable to use as atherapeutic agent only one of a pair of enantiomers, or only one of aplurality of diastereoisomers. Accordingly, the invention providescompositions containing a compound of the formula (I) having one or morechiral centres, wherein at least 55% (e.g. at least 60%, 65%, 70%, 75%,80%, 85%, 90% or 95%) of the compound of the formula (I) is present as asingle optical isomer (e.g. enantiomer or diastereoisomer). In onegeneral embodiment, 99% or more (e.g. substantially all) of the totalamount of the compound of the formula (I) may be present as a singleoptical isomer (e.g. enantiomer or diastereoisomer). When a specificisomeric form is identified (e.g. S configuration, or E isomer), thismeans that said isomeric form is substantially free of the otherisomer(s), i.e. said isomeric form is present in at least 55%, 60%, 65%,70%, 75%, 80%, 85%, 90%, 95%, 99% or more (e.g. substantially all) ofthe total amount of the compound of the invention.

Whenever, hereinbefore or hereinafter, compounds include the followingbond

, this indicates that the compound is a single stereoisomer with unknownconfiguration or a mixture of stereoisomers.

The compounds of the invention include compounds with one or moreisotopic substitutions, and a reference to a particular element includeswithin its scope all isotopes of the element. For example, a referenceto hydrogen includes within its scope ¹H, ²H (D), and ³H (T). Similarly,references to carbon and oxygen include within their scope respectively¹²C, ¹³C and ¹⁴C and ¹⁶O and ¹⁸O. The isotopes may be radioactive ornon-radioactive. In one embodiment of the invention, the compoundscontain no radioactive isotopes. Such compounds are preferred fortherapeutic use. In another embodiment, however, the compound maycontain one or more radioisotopes. Compounds containing suchradioisotopes may be useful in a diagnostic context.

Esters such as carboxylic acid esters and acyloxy esters of thecompounds of formula (I) bearing a carboxylic acid group or a hydroxylgroup are also embraced by formula (I). In one embodiment of theinvention, formula (I) includes within its scope esters of compounds ofthe formula (I) bearing a hydroxyl group. In another embodiment of theinvention, formula (I) does not include within its scope esters ofcompounds of the formula (I) bearing a hydroxyl group. Examples ofacyloxy (reverse ester) groups are represented by —OC(═O)R, wherein R isan acyloxy substituent, for example, a C₁₋₇ alkyl group, a C₃₋₂₀heterocyclyl group, or a C₅₋₂₀ aryl group, preferably a C₁₋₇ alkylgroup. Particular examples of acyloxy groups include, but are notlimited to, —OC(═O)CH₃ (acetoxy), —OC(═O)CH₂CH₃, —OC(═O)C(CH₃)₃,—OC(═O)Ph, and —OC(═O)CH₂Ph.

For example, some prodrugs are esters of the active compound (e.g., aphysiologically acceptable metabolically labile ester). By “prodrugs” ismeant for example any compound that is converted in vivo into abiologically active compound of the formula (I). During metabolism, theester group is cleaved to yield the active drug. Such esters may beformed by esterification, for example, of any of the hydroxyl groups inthe parent compound, with, where appropriate, prior protection of anyother reactive groups present in the parent compound, followed bydeprotection if required.

Examples of such metabolically labile esters include C₁₋₆ aminoalkyl[e.g., aminoethyl; 2-(N,N-diethylamino)ethyl; 2-(4-morpholino)ethyl);and acyloxy-C₁₋₇alkyl [e.g., acyloxymethyl; acyloxyethyl;pivaloyloxymethyl; acetoxymethyl; 1-acetoxyethyl;1-(1-methoxy-1-methyl)ethyl-carbonyloxyethyl; 1-(benzoyloxy)ethyl;isopropoxy-carbonyloxymethyl; 1-isopropoxy-carbonyloxyethyl;cyclohexyl-carbonyloxymethyl; 1-cyclohexyl-carbonyloxyethyl;cyclohexyloxy-carbonyloxymethyl; 1-cyclohexyloxy-carbonyloxyethyl;(4-tetrahydropyranyloxy) carbonyloxymethyl;1-(4-tetrahydropyranyloxy)carbonyloxyethyl;(4-tetrahydropyranyl)carbonyloxymethyl; and1-(4-tetrahydropyranyl)carbonyloxyethyl]. Also, some prodrugs areactivated enzymatically to yield the active compound, or a compoundwhich, upon further chemical reaction, yields the active compound (forexample, as in antigen-directed enzyme pro-drug therapy (ADEPT),gene-directed enzyme pro-drug therapy (GDEPT) and ligand-directed enzymepro-drug therapy (LIDEPT) etc.). For example, the prodrug may be a sugarderivative or other glycoside conjugate, or may be an amino acid esterderivative.

Protein Tyrosine Kinases (PTK)

The compounds of the invention described herein inhibit or modulate theactivity of certain tyrosine kinases, and thus the compounds will beuseful in the treatment or prophylaxis, in particular the treatment, ofdisease states or conditions mediated by those tyrosine kinases, inparticular FGFR.

FGFR

The fibroblast growth factor (FGF) family of protein tyrosine kinase(PTK) receptors regulates a diverse array of physiologic functionsincluding mitogenesis, wound healing, cell differentiation andangiogenesis, and development. Both normal and malignant cell growth aswell as proliferation are affected by changes in local concentration ofFGFs, extracellular signalling molecules which act as autocrine as wellas paracrine factors. Autocrine FGF signalling may be particularlyimportant in the progression of steroid hormone-dependent cancers to ahormone independent state. FGFs and their receptors are expressed atincreased levels in several tissues and cell lines and overexpression isbelieved to contribute to the malignant phenotype. Furthermore, a numberof oncogenes are homologues of genes encoding growth factor receptors,and there is a potential for aberrant activation of FGF-dependentsignalling in human pancreatic cancer (Knights et al., Pharmacology andTherapeutics 2010 125:1 (105-117); Korc M. et al Current Cancer DrugTargets 2009 9:5 (639-651)).

The two prototypic members are acidic fibroblast growth factor (aFGF orFGF1) and basic fibroblast growth factor (bFGF or FGF2), and to date, atleast twenty distinct FGF family members have been identified. Thecellular response to FGFs is transmitted via four types of high affinitytransmembrane protein tyrosine-kinase fibroblast growth factor receptors(FGFR) numbered 1 to 4 (FGFR1 to FGFR4).

Disruption of the FGFR1 pathway should affect tumor cell proliferationsince this kinase is activated in many tumor types in addition toproliferating endothelial cells. The over-expression and activation ofFGFR1 in tumor-associated vasculature has suggested a role for thesemolecules in tumor angiogenesis.

A recent study has shown a link between FGFR1 expression andtumorigenicity in Classic Lobular Carcinomas (CLC). CLCs account for10-15% of all breast cancers and, in general, lack p53 and Her2expression whilst retaining expression of the oestrogen receptor. A geneamplification of 8p12-p11.2 was demonstrated in ˜50% of CLC cases andthis was shown to be linked with an increased expression of FGFR1.Preliminary studies with siRNA directed against FGFR1, or a smallmolecule inhibitor of the receptor, showed cell lines harbouring thisamplification to be particularly sensitive to inhibition of thissignalling pathway. Rhabdomyosarcoma (RMS) is the most common pediatricsoft tissue sarcoma likely results from abnormal proliferation anddifferentiation during skeletal myogenesis. FGFR1 is over-expressed inprimary rhabdomyosarcoma tumors and is associated with hypomethylationof a 5′ CpG island and abnormal expression of the AKT1, NOG, and BMP4genes. FGFR1 has also been linked to squamous lung cancer, colorectalcancer, glioblastoma, astrocytomas, prostate cancer, small cell lungcancer, melanoma, head and neck cancer, thyroid cancer, uterine cancer.

Fibroblast growth factor receptor 2 has high affinity for the acidicand/or basic fibroblast growth factors, as well as the keratinocytegrowth factor ligands. Fibroblast growth factor receptor 2 alsopropagates the potent osteogenic effects of FGFs during osteoblastgrowth and differentiation. Mutations in fibroblast growth factorreceptor 2, leading to complex functional alterations, were shown toinduce abnormal ossification of cranial sutures (craniosynostosis),implying a major role of FGFR signalling in intramembranous boneformation. For example, in Apert (AP) syndrome, characterized bypremature cranial suture ossification, most cases are associated withpoint mutations engendering gain-of-function in fibroblast growth factorreceptor 2. In addition, mutation screening in patients with syndromiccraniosynostoses indicates that a number of recurrent FGFR2 mutationsaccounts for severe forms of Pfeiffer syndrome. Particular mutations ofFGFR2 include W290C, D321A, Y340C, C342R, C342S, C342W, N549H, K641R inFGFR2.

Several severe abnormalities in human skeletal development, includingApert, Crouzon, Jackson-Weiss, Beare-Stevenson cutis gyrata, andPfeiffer syndromes are associated with the occurrence of mutations infibroblast growth factor receptor 2. Most, if not all, cases of PfeifferSyndrome (PS) are also caused by de novo mutation of the fibroblastgrowth factor receptor 2 gene, and it was recently shown that mutationsin fibroblast growth factor receptor 2 break one of the cardinal rulesgoverning ligand specificity. Namely, two mutant splice forms offibroblast growth factor receptor, FGFR2c and FGFR2b, have acquired theability to bind to and be activated by atypical FGF ligands. This lossof ligand specificity leads to aberrant signalling and suggests that thesevere phenotypes of these disease syndromes result from ectopicligand-dependent activation of fibroblast growth factor receptor 2.

Genetic aberrations of the FGFR3 receptor tyrosine kinase such aschromosomal translocations or point mutations result in ectopicallyexpressed or deregulated, constitutively active, FGFR3 receptors. Suchabnormalities are linked to a subset of multiple myelomas and inbladder, hepatocellular, oral squamous cell carcinoma and cervicalcarcinomas. Accordingly, FGFR3 inhibitors would be useful in thetreatment of multiple myeloma, bladder and cervical carcinomas. FGFR3 isalso over-expressed in bladder cancer, in particular invasive bladdercancer. FGFR3 is frequently activated by mutation in urothelialcarcinoma (UC). Increased expression was associated with mutation (85%of mutant tumors showed high-level expression) but also 42% of tumorswith no detectable mutation showed over-expression, including manymuscle-invasive tumors. FGFR3 is also linked to endometrial and thyroidcancer.

Over expression of FGFR4 has been linked to poor prognosis in bothprostate and thyroid carcinomas. In addition a germline polymorphism(Gly388Arg) is associated with increased incidence of lung, breast,colon, liver (HCC) and prostate cancers. In addition, a truncated formof FGFR4 (including the kinase domain) has also been found to be presentin 40% of pituitary tumours but not present in normal tissue. FGFR4overexpression has been observed in liver, colon and lung tumours. FGFR4has been implicated in colorectal and liver cancer where expression ofits ligand FGF19 is frequently elevated. FGFR4 is also linked toastrocytomas, rhabdomyosarcoma.

Fibrotic conditions are a major medical problem resulting from abnormalor excessive deposition of fibrous tissue. This occurs in many diseases,including liver cirrhosis, glomerulonephritis, pulmonary fibrosis,systemic fibrosis, rheumatoid arthritis, as well as the natural processof wound healing. The mechanisms of pathological fibrosis are not fullyunderstood but are thought to result from the actions of variouscytokines (including tumor necrosis factor (TNF), fibroblast growthfactors (FGF's), platelet derived growth factor (PDGF) and transforminggrowth factor beta. (TGFβ) involved in the proliferation of fibroblastsand the deposition of extracellular matrix proteins (including collagenand fibronectin). This results in alteration of tissue structure andfunction and subsequent pathology.

A number of preclinical studies have demonstrated the up-regulation offibroblast growth factors in preclinical models of lung fibrosis. TGFβ1and PDGF have been reported to be involved in the fibrogenic process andfurther published work suggests the elevation of FGF's and consequentincrease in fibroblast proliferation, may be in response to elevatedTGFβ1. The potential therapeutic benefit of targeting the fibroticmechanism in conditions such as idiopathic pulmonary fibrosis (IPF) issuggested by the reported clinical effect of the anti-fibrotic agentpirfenidone. Idiopathic pulmonary fibrosis (also referred to asCryptogenic fibrosing alveolitis) is a progressive condition involvingscarring of the lung. Gradually, the air sacs of the lungs becomereplaced by fibrotic tissue, which becomes thicker, causing anirreversible loss of the tissue's ability to transfer oxygen into thebloodstream. The symptoms of the condition include shortness of breath,chronic dry coughing, fatigue, chest pain and loss of appetite resultingin rapid weight loss. The condition is extremely serious withapproximately 50% mortality after 5 years.

As such, the compounds which inhibit FGFR will be useful in providing ameans of preventing the growth or inducing apoptosis in tumours,particularly by inhibiting angiogenesis. It is therefore anticipatedthat the compounds will prove useful in treating or preventingproliferative disorders such as cancers. In particular tumours withactivating mutants of receptor tyrosine kinases (RTK) or upregulation ofreceptor tyrosine kinases may be particularly sensitive to theinhibitors. Patients with activating mutants of any of the isoforms ofthe specific RTKs discussed herein may also find treatment with RTKinhibitors particularly beneficial, for instance patients with tumors,e.g. bladder or brain tumors, with FGFR3-TACC3 translocation.

Vascular Endothelial Growth Factor Receptor (VEGFR)

Chronic proliferative diseases are often accompanied by profoundangiogenesis, which can contribute to or maintain an inflammatory and/orproliferative state, or which leads to tissue destruction through theinvasive proliferation of blood vessels.

Angiogenesis is generally used to describe the development of new orreplacement blood vessels, or neovascularisation. It is a necessary andphysiological normal process by which vasculature is established in theembryo. Angiogenesis does not occur, in general, in most normal adulttissues, exceptions being sites of ovulation, menses and wound healing.Many diseases, however, are characterized by persistent and unregulatedangiogenesis. For instance, in arthritis, new capillary blood vesselsinvade the joint and destroy cartilage. In diabetes (and in manydifferent eye diseases), new vessels invade the macula or retina orother ocular structures, and may cause blindness. The process ofatherosclerosis has been linked to angiogenesis. Tumor growth andmetastasis have been found to be angiogenesis-dependent.

The recognition of the involvement of angiogenesis in major diseases hasbeen accompanied by research to identify and develop inhibitors ofangiogenesis. These inhibitors are generally classified in response todiscrete targets in the angiogenesis cascade, such as activation ofendothelial cells by an angiogenic signal; synthesis and release ofdegradative enzymes; endothelial cell migration; proliferation ofendothelial cells; and formation of capillary tubules. Therefore,angiogenesis occurs in many stages and attempts are underway to discoverand develop compounds that work to block angiogenesis at these variousstages.

There are publications that teach that inhibitors of angiogenesis,working by diverse mechanisms, are beneficial in diseases such as cancerand metastasis, ocular diseases, arthritis and hemangioma.

Vascular endothelial growth factor (VEGF), a polypeptide, is mitogenicfor endothelial cells in vitro and stimulates angiogenic responses invivo. VEGF has also been linked to inappropriate angiogenesis. VEGFR(s)are protein tyrosine kinases (PTKs). PTKs catalyze the phosphorylationof specific tyrosine residues in proteins involved in cell function thusregulating cell growth, survival and differentiation.

Three PTK receptors for VEGF have been identified: VEGFR-1 (Flt-1);VEGFR-2 (Flk-1 or KDR) and VEGFR-3 (Flt-4). These receptors are involvedin angiogenesis and participate in signal transduction. Of particularinterest is VEGFR-2, which is a transmembrane receptor PTK expressedprimarily in endothelial cells. Activation of VEGFR-2 by VEGF is acritical step in the signal transduction pathway that initiates tumourangiogenesis. VEGF expression may be constitutive to tumour cells andcan also be upregulated in response to certain stimuli. One such stimuliis hypoxia, where VEGF expression is upregulated in both tumour andassociated host tissues. The VEGF ligand activates VEGFR-2 by bindingwith its extracellular VEGF binding site. This leads to receptordimerization of VEGFRs and autophosphorylation of tyrosine residues atthe intracellular kinase domain of VEGFR-2. The kinase domain operatesto transfer a phosphate from ATP to the tyrosine residues, thusproviding binding sites for signalling proteins downstream of VEGFR-2leading ultimately to initiation of angiogenesis.

Inhibition at the kinase domain binding site of VEGFR-2 would blockphosphorylation of tyrosine residues and serve to disrupt initiation ofangiogenesis.

Angiogenesis is a physiologic process of new blood vessel formationmediated by various cytokines called angiogenic factors. Although itspotential pathophysiologic role in solid tumors has been extensivelystudied for more than 3 decades, enhancement of angiogenesis in chroniclymphocytic leukemia (CLL) and other malignant hematological disordershas been recognized more recently. An increased level of angiogenesishas been documented by various experimental methods both in bone marrowand lymph nodes of patients with CLL. Although the role of angiogenesisin the pathophysiology of this disease remains to be fully elucidated,experimental data suggest that several angiogenic factors play a role inthe disease progression. Biologic markers of angiogenesis were alsoshown to be of prognostic relevance in CLL. This indicates that VEGFRinhibitors may also be of benefit for patients with leukemia's such asCLL.

In order for a tumour mass to get beyond a critical size, it mustdevelop an associated vasculature. It has been proposed that targeting atumor vasculature would limit tumor expansion and could be a usefulcancer therapy. Observations of tumor growth have indicated that smalltumour masses can persist in a tissue without any tumour-specificvasculature. The growth arrest of nonvascularized tumors has beenattributed to the effects of hypoxia at the center of the tumor. Morerecently, a variety of proangiogenic and antiangiogenic factors havebeen identified and have led to the concept of the “angiogenic switch,”a process in which disruption of the normal ratio of angiogenic stimuliand inhibitors in a tumor mass allows for autonomous vascularization.The angiogenic switch appears to be governed by the same geneticalterations that drive malignant conversion: the activation of oncogenesand the loss of tumour suppressor genes. Several growth factors act aspositive regulators of angiogenesis. Foremost among these are vascularendothelial growth factor (VEGF), basic fibroblast growth factor (bFGF),and angiogenin. Proteins such as thrombospondin (Tsp-1), angiostatin,and endostatin function as negative regulators of angiogenesis.

Inhibition of VEGFR2 but not VEGFR1 markedly disrupts angiogenicswitching, persistent angiogenesis, and initial tumor growth in a mousemodel. In late-stage tumors, phenotypic resistance to VEGFR2 blockadeemerged, as tumors regrew during treatment after an initial period ofgrowth suppression. This resistance to VEGF blockade involvesreactivation of tumour angiogenesis, independent of VEGF and associatedwith hypoxia-mediated induction of other proangiogenic factors,including members of the FGF family. These other proangiogenic signalsare functionally implicated in the revascularization and regrowth oftumours in the evasion phase, as FGF blockade impairs progression in theface of VEGF inhibition.

There is evidence for normalization of glioblastoma blood vessels inpatients treated with a pan-VEGF receptor tyrosine kinase inhibitor,AZD2171, in a phase 2 study. MRI determination of vessel normalizationin combination with circulating biomarkers provides for an effectivemeans to assess response to antiangiogenic agents.

PDGFR

A malignant tumour is the product of uncontrolled cell proliferation.Cell growth is controlled by a delicate balance between growth-promotingand growth-inhibiting factors. In normal tissue the production andactivity of these factors results in differentiated cells growing in acontrolled and regulated manner that maintains the normal integrity andfunctioning of the organ. The malignant cell has evaded this control;the natural balance is disturbed (via a variety of mechanisms) andunregulated, aberrant cell growth occurs. A growth factor of importancein tumour development is the platelet-derived growth factor (PDGF) thatcomprises a family of peptide growth factors that signal through cellsurface tyrosine kinase receptors (PDGFR) and stimulate various cellularfunctions including growth, proliferation, and differentiation.

Advantages of a Selective Inhibitor

Development of FGFR kinase inhibitors with a differentiated selectivityprofile provides a new opportunity to use these targeted agents inpatient sub-groups whose disease is driven by FGFR deregulation.Compounds that exhibit reduced inhibitory action on additional kinases,particularly VEGFR2 and PDGFR-beta, offer the opportunity to have adifferentiated side-effect or toxicity profile and as such allow for amore effective treatment of these indications. Inhibitors of VEGFR2 andPDGFR-beta are associated with toxicities such as hypertension or oedemarespectively. In the case of VEGFR2 inhibitors this hypertensive effectis often dose limiting, may be contraindicated in certain patientpopulations and requires clinical management.

Biological Activity and Therapeutic Uses

The compounds of the invention, and subgroups thereof, have fibroblastgrowth factor receptor (FGFR) inhibiting or modulating activity and/orvascular endothelial growth factor receptor (VEGFR) inhibiting ormodulating activity, and/or platelet derived growth factor receptor(PDGFR) inhibiting or modulating activity, and which will be useful inpreventing or treating disease states or conditions described herein. Inaddition the compounds of the invention, and subgroups thereof, will beuseful in preventing or treating diseases or condition mediated by thekinases. References to the preventing or prophylaxis or treatment of adisease state or condition such as cancer include within their scopealleviating or reducing the incidence of cancer.

As used herein, the term “modulation”, as applied to the activity of akinase, is intended to define a change in the level of biologicalactivity of the protein kinase. Thus, modulation encompassesphysiological changes which effect an increase or decrease in therelevant protein kinase activity. In the latter case, the modulation maybe described as “inhibition”. The modulation may arise directly orindirectly, and may be mediated by any mechanism and at anyphysiological level, including for example at the level of geneexpression (including for example transcription, translation and/orpost-translational modification), at the level of expression of genesencoding regulatory elements which act directly or indirectly on thelevels of kinase activity. Thus, modulation may implyelevated/suppressed expression or over- or under-expression of a kinase,including gene amplification (i.e. multiple gene copies) and/orincreased or decreased expression by a transcriptional effect, as wellas hyper-(or hypo-)activity and (de)activation of the protein kinase(s)(including (de)activation) by mutation(s). The terms “modulated”,“modulating” and “modulate” are to be interpreted accordingly.

As used herein, the term “mediated”, as used e.g. in conjunction with akinase as described herein (and applied for example to variousphysiological processes, diseases, states, conditions, therapies,treatments or interventions) is intended to operate limitatively so thatthe various processes, diseases, states, conditions, treatments andinterventions to which the term is applied are those in which the kinaseplays a biological role. In cases where the term is applied to adisease, state or condition, the biological role played by a kinase maybe direct or indirect and may be necessary and/or sufficient for themanifestation of the symptoms of the disease, state or condition (or itsaetiology or progression). Thus, kinase activity (and in particularaberrant levels of kinase activity, e.g. kinase over-expression) neednot necessarily be the proximal cause of the disease, state orcondition: rather, it is contemplated that the kinase mediated diseases,states or conditions include those having multifactorial aetiologies andcomplex progressions in which the kinase in question is only partiallyinvolved. In cases where the term is applied to treatment, prophylaxisor intervention, the role played by the kinase may be direct or indirectand may be necessary and/or sufficient for the operation of thetreatment, prophylaxis or outcome of the intervention. Thus, a diseasestate or condition mediated by a kinase includes the development ofresistance to any particular cancer drug or treatment.

Thus, for example, the compounds of the invention may be useful inalleviating or reducing the incidence of cancer.

More particularly, the compounds of the formulae (I) and sub-groupsthereof are inhibitors of FGFRs. For example, compounds of the inventionhave activity against FGFR1, FGFR2, FGFR3, and/or FGFR4, and inparticular FGFRs selected from FGFR1, FGFR2 and FGFR3; or in particularthe compounds of formula (I) and sub-groups thereof are inhibitors ofFGFR4.

Preferred compounds are compounds that inhibit one or more FGFR selectedfrom FGFR1, FGFR2, FGFR3, and FGFR4. Preferred compounds of theinvention are those having IC₅₀ values of less than 0.1 μM.

Compounds of the invention also have activity against VEGFR.

In addition many of the compounds of the invention exhibit selectivityfor the FGFR 1, 2, and/or 3, and/or 4 compared to VEGFR (in particularVEGFR2) and/or PDGFR and such compounds represent one preferredembodiment of the invention. In particular, the compounds exhibitselectivity over VEGFR2. For example, many compounds of the inventionhave IC₅₀ values against FGFR1, 2 and/or 3 and/or 4 that are between atenth and a hundredth of the IC₅₀ against VEGFR (in particular VEGFR2)and/or PDGFR B. In particular preferred compounds of the invention haveat least 10 times greater activity against or inhibition of FGFR inparticular FGFR1, FGFR2, FGFR3 and/or FGFR4 than VEGFR2. More preferablythe compounds of the invention have at least 100 times greater activityagainst or inhibition of FGFR in particular FGFR1, FGFR2, FGFR3 and/orFGFR4 than VEGFR2. This can be determined using the methods describedherein.

As a consequence of their activity in modulating or inhibiting FGFR,and/or VEGFR kinases, the compounds will be useful in providing a meansof preventing the growth or inducing apoptosis of neoplasias,particularly by inhibiting angiogenesis. It is therefore anticipatedthat the compounds will prove useful in treating or preventingproliferative disorders such as cancers. In addition, the compounds ofthe invention could be useful in the treatment of diseases in whichthere is a disorder of proliferation, apoptosis or differentiation.

In particular tumours with activating mutants of VEGFR or upregulationof VEGFR and patients with elevated levels of serum lactatedehydrogenase may be particularly sensitive to the compounds of theinvention. Patients with activating mutants of any of the isoforms ofthe specific RTKs discussed herein may also find treatment with thecompounds of the invention particularly beneficial. For example, VEGFRoverexpression in acute leukemia cells where the clonal progenitor mayexpress VEGFR. Also, particular tumours with activating mutants orupregulation or overexpression of any of the isoforms of FGFR such asFGFR1, FGFR2 or FGFR3 or FGFR4 may be particularly sensitive to thecompounds of the invention and thus patients as discussed herein withsuch particular tumours may also find treatment with the compounds ofthe invention particularly beneficial. It may be preferred that thetreatment is related to or directed at a mutated form of one of thereceptor tyrosine kinases, such as discussed herein. Diagnosis oftumours with such mutations could be performed using techniques known toa person skilled in the art and as described herein such as RTPCR andFISH.

Examples of cancers which may be treated (or inhibited) include, but arenot limited to, a carcinoma, for example a carcinoma of the bladder,breast, colon (e.g. colorectal carcinomas such as colon adenocarcinomaand colon adenoma), kidney, urothelial, uterus, epidermis, liver, lung(for example adenocarcinoma, small cell lung cancer and non-small celllung carcinomas, squamous lung cancer), oesophagus, head and neck, gallbladder, ovary, pancreas (e.g. exocrine pancreatic carcinoma), stomach,gastrointestinal (also known as gastric) cancer (e.g. gastrointestinalstromal tumours), cervix, endometrium, thyroid, prostate, or skin (forexample squamous cell carcinoma or dermatofibrosarcoma protuberans);pituitary cancer, a hematopoietic tumour of lymphoid lineage, forexample leukemia, acute lymphocytic leukemia, chronic lymphocyticleukemia, B-cell lymphoma (e.g. diffuse large B-cell lymphoma), T-celllymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, hairy celllymphoma, or Burkett's lymphoma; a hematopoietic tumour of myeloidlineage, for example leukemias, acute and chronic myelogenous leukemias,chronic myelomonocytic leukemia (CMML), myeloproliferative disorder,myeloproliferative syndrome, myelodysplastic syndrome, or promyelocyticleukemia; multiple myeloma; thyroid follicular cancer; hepatocellularcancer, a tumour of mesenchymal origin (e.g. Ewing's sarcoma), forexample fibrosarcoma or rhabdomyosarcoma; a tumour of the central orperipheral nervous system, for example astrocytoma, neuroblastoma,glioma (such as glioblastoma multiforme) or schwannoma; melanoma;seminoma; teratocarcinoma; osteosarcoma; xeroderma pigmentosum;keratoctanthoma; thyroid follicular cancer; or Kaposi's sarcoma. Inparticular, squamous lung cancer, breast cancer, colorectal cancer,glioblastoma, astrocytomas, prostate cancer, small cell lung cancer,melanoma, head and neck cancer, thyroid cancer, uterine cancer, gastriccancer, hepatocellular cancer, cervix cancer, multiple myeloma, bladdercancer, endometrial cancer, urothelial cancer, colon cancer,rhabdomyosarcoma, pituitary gland cancer.

Examples of cancers which may be treated (or inhibited) include, but arenot limited to, bladder cancer, urothelial cancer, metastatic urothelialcancer, surgically unresectable urothelial cancer, breast cancer,glioblastoma, lung cancer, non small cell lung cancer, squamous celllung cancer, adenocarcinoma of the lung, pulmonary adenocarcinoma, smallcell lung cancer, ovarian cancer, endometrial cancer, cervical cancer,soft tissue sarcoma, head and neck squamous cell carcinoma, gastriccancer, oesophageal cancer, squamous cell carcinoma of the oesophagus,adenocarcinoma of the oesophagus, cholangiocarcinoma, hepatocellularcarcinoma.

Certain cancers are resistant to treatment with particular drugs. Thiscan be due to the type of the tumour or can arise due to treatment withthe compound. In this regard, references to multiple myeloma includesbortezomib sensitive multiple myeloma or refractory multiple myeloma.Similarly, references to chronic myelogenous leukemia includes imitanibsensitive chronic myelogenous leukemia and refractory chronicmyelogenous leukemia. Chronic myelogenous leukemia is also known aschronic myeloid leukemia, chronic granulocytic leukemia or CML.Likewise, acute myelogenous leukemia, is also called acute myeloblasticleukemia, acute granulocytic leukemia, acute nonlymphocytic leukaemia orAML.

The compounds of the invention can also be used in the treatment ofhematopoetic diseases of abnormal cell proliferation whetherpre-malignant or stable such as myeloproliferative diseases.Myeloproliferative diseases (“MPD” s) are a group of diseases of thebone marrow in which excess cells are produced. They are related to, andmay evolve into, myelodysplastic syndrome. Myeloproliferative diseasesinclude polycythemia vera, essential thrombocythemia and primarymyelofibrosis. A further haematological disorder is hypereosinophilicsyndrome. T-cell lymphoproliferative diseases include those derived fromnatural Killer cells.

In addition the compounds of the invention can be used togastrointestinal (also known as gastric) cancer e.g. gastrointestinalstromal tumours. Gastrointestinal cancer refers to malignant conditionsof the gastrointestinal tract, including the esophagus, stomach, liver,biliary system, pancreas, bowels, and anus.

Thus, in the pharmaceutical compositions, uses or methods of thisinvention for treating a disease or condition comprising abnormal cellgrowth, the disease or condition comprising abnormal cell growth in oneembodiment is a cancer.

Particular subsets of cancers include multiple myeloma, bladder,cervical, prostate and thyroid carcinomas, lung, breast, and coloncancers.

A further subset of cancers includes multiple myeloma, bladder,hepatocellular, oral squamous cell carcinoma and cervical carcinomas.

The compound of the invention, having FGFR such as FGFR1 inhibitoryactivity, may be particularly useful in the treatment or prevention ofbreast cancer in particular Classic Lobular Carcinomas (CLC).

As the compounds of the invention have FGFR4 activity they will also beuseful in the treatment of prostate or pituitary cancers, or they willbe useful in the treatment of breast cancer, lung cancer, prostatecancer, liver cancer (HCC) or lung cancer.

In particular the compounds of the invention as FGFR inhibitors, areuseful in the treatment of multiple myeloma, myeloproliferatoivedisorders, endometrial cancer, prostate cancer, bladder cancer, lungcancer, ovarian cancer, breast cancer, gastric cancer, colorectalcancer, and oral squamous cell carcinoma.

Further subsets of cancer are multiple myeloma, endometrial cancer,bladder cancer, cervical cancer, prostate cancer, lung cancer, breastcancer, colorectal cancer and thyroid carcinomas.

In particular the compounds of the invention are useful in the treatmentof multiple myeloma (in particular multiple myeloma with t(4; 14)translocation or overexpressing FGFR3), prostate cancer (hormonerefractory prostrate carcinomas), endometrial cancer (in particularendometrial tumours with activating mutations in FGFR2) and breastcancer (in particular lobular breast cancer).

In particular the compounds are useful in the treatment of lobularcarcinomas such as CLC (Classic lobular carcinoma).

As the compounds have activity against FGFR3 they will be useful in thetreatment of multiple myeloma and bladder cancer.

In particular, the compounds have activity against tumours withFGFR3-TACC3 translocation, in particular bladder or brain tumours withFGFR3-TACC3 translocation.

In particular the compounds are useful for the treatment of t(4; 14)translocation positive multiple myeloma.

In one embodiment the compounds may be useful for the treatment ofsarcoma. In one embodiment the compounds may be useful for the treatmentof lung cancer, e.g. squamous cell carcinoma.

As the compounds have activity against FGFR2 they will be useful in thetreatment of endometrial, ovarian, gastric, hepatocellular, uterine,cervix and colorectal cancers. FGFR2 is also overexpressed in epithelialovarian cancer, therefore the compounds of the invention may bespecifically useful in treating ovarian cancer such as epithelialovarian cancer.

In one embodiment, the compounds may be useful for the treatment of lungcancer, in particular NSCLC, squamous cell carcinoma, liver cancer,kidney cancer, breast cancer, colon cancer, colorectal cancer, prostatecancer.

Compounds of the invention may also be useful in the treatment oftumours pre-treated with VEGFR2 inhibitor or VEGFR2 antibody (e.g.Avastin).

In particular the compounds of the invention may be useful in thetreatment of VEGFR2-resistant tumours. VEGFR2 inhibitors and antibodiesare used in the treatment of thyroid and renal cell carcinomas,therefore the compounds of the invention may be useful in the treatmentof VEGFR2-resistant thyroid and renal cell carcinomas.

The cancers may be cancers which are sensitive to inhibition of any oneor more FGFRs selected from FGFR1, FGFR2, FGFR3, FGFR4, for example, oneor more FGFRs selected from FGFR1, FGFR2 or FGFR3.

Whether or not a particular cancer is one which is sensitive toinhibition of FGFR or VEGFR signalling may be determined by means of acell growth assay as set out below or by a method as set out in thesection headed “Methods of Diagnosis”.

The compounds of the invention, and in particular those compounds havingFGFR, or VEGFR inhibitory activity, may be particularly useful in thetreatment or prevention of cancers of a type associated with orcharacterised by the presence of elevated levels of FGFR, or VEGFR, forexample the cancers referred to in this context in the introductorysection of this application.

The compounds of the present invention may be useful for the treatmentof the adult population. The compounds of the present invention may beuseful for the treatment of the pediatric population.

It has been discovered that some FGFR inhibitors can be used incombination with other anticancer agents. For example, it may bebeneficial to combine an inhibitor that induces apoptosis with anotheragent which acts via a different mechanism to regulate cell growth thustreating two of the characteristic features of cancer development.Examples of such combinations are set out below.

The compounds of the invention may be useful in treating otherconditions which result from disorders in proliferation such as type IIor non-insulin dependent diabetes mellitus, autoimmune diseases, headtrauma, stroke, epilepsy, neurodegenerative diseases such asAlzheimer's, motor neurone disease, progressive supranuclear palsy,corticobasal degeneration and Pick's disease for example autoimmunediseases and neurodegenerative diseases.

One sub-group of disease states and conditions that the compounds of theinvention may be useful consists of inflammatory diseases,cardiovascular diseases and wound healing.

FGFR, and VEGFR are also known to play a role in apoptosis,angiogenesis, proliferation, differentiation and transcription andtherefore the compounds of the invention could also be useful in thetreatment of the following diseases other than cancer; chronicinflammatory diseases, for example systemic lupus erythematosus,autoimmune mediated glomerulonephritis, rheumatoid arthritis, psoriasis,inflammatory bowel disease, autoimmune diabetes mellitus, Eczemahypersensitivity reactions, asthma, COPD, rhinitis, and upperrespiratory tract disease; cardiovascular diseases for example cardiachypertrophy, restenosis, atherosclerosis; neurodegenerative disorders,for example Alzheimer's disease, AIDS-related dementia, Parkinson'sdisease, amyotropic lateral sclerosis, retinitis pigmentosa, spinalmuscular atropy and cerebellar degeneration; glomerulonephritis;myelodysplastic syndromes, ischemic injury associated myocardialinfarctions, stroke and reperfusion injury, arrhythmia, atherosclerosis,toxin-induced or alcohol related liver diseases, haematologicaldiseases, for example, chronic anemia and aplastic anemia; degenerativediseases of the musculoskeletal system, for example, osteoporosis andarthritis, aspirin-sensitive rhinosinusitis, cystic fibrosis, multiplesclerosis, kidney diseases and cancer pain.

In addition, mutations of FGFR2 are associated with several severeabnormalities in human skeletal development and thus the compounds ofinvention could be useful in the treatment of abnormalities in humanskeletal development, including abnormal ossification of cranial sutures(craniosynostosis), Apert (AP) syndrome, Crouzon syndrome, Jackson-Weisssyndrome, Beare-Stevenson cutis gyrate syndrome, and Pfeiffer syndrome.

The compound of the invention, having FGFR such as FGFR2 or FGFR3inhibitory activity, may be particularly useful in the treatment orprevention of the skeletal diseases. Particular skeletal diseases areachondroplasia or thanatophoric dwarfism (also known as thanatophoricdysplasia).

The compound of the invention, having FGFR such as FGFR1, FGFR2 or FGFR3inhibitory activity, may be particularly useful in the treatment orprevention in pathologies in which progressive fibrosis is a symptom.Fibrotic conditions in which the compounds of the inventions may beuseful in the treatment of include diseases exhibiting abnormal orexcessive deposition of fibrous tissue for example in liver cirrhosis,glomerulonephritis, pulmonary fibrosis, systemic fibrosis, rheumatoidarthritis, as well as the natural process of wound healing. Inparticular the compounds of the inventions may also be useful in thetreatment of lung fibrosis in particular in idiopathic pulmonaryfibrosis.

The over-expression and activation of FGFR and VEGFR in tumor-associatedvasculature has also suggested a role for compounds of the invention inpreventing and disrupting initiation of tumor angiogenesis. Inparticular the compounds of the invention may be useful in the treatmentof cancer, metastasis, leukemia's such as CLL, ocular diseases such asage-related macular degeneration in particular wet form of age-relatedmacular degeneration, ischemic proliferative retinopathies such asretinopathy of prematurity (ROP) and diabetic retinopathy, rheumatoidarthritis and hemangioma.

The activity of the compounds of the invention as inhibitors of FGFR1-4,VEGFR and/or PDGFR AB can be measured using the assays set forth in theexamples below and the level of activity exhibited by a given compoundcan be defined in terms of the IC₅₀ value. Preferred compounds of thepresent invention are compounds having an IC₅₀ value of less than 1 μM,more preferably less than 0.1 μM.

The invention provides compounds that have FGFR inhibiting or modulatingactivity, and which may be useful in preventing or treating diseasestates or conditions mediated by FGFR kinases.

In one embodiment, there is provided a compound as defined herein foruse in therapy, for use as a medicine. In a further embodiment, there isprovided a compound as defined herein for use in the prophylaxis ortreatment, in particular in the treatment, of a disease state orcondition mediated by a FGFR kinase.

Thus, for example, the compounds of the invention may be useful inalleviating or reducing the incidence of cancer. Therefore, in a furtherembodiment, there is provided a compound as defined herein for use inthe prophylaxis or treatment, in particular the treatment, of cancer. Inone embodiment, the compound as defined herein is for use in theprophylaxis or treatment of FGFR-dependent cancer. In one embodiment,the compound as defined herein is for use in the prophylaxis ortreatment of cancer mediated by FGFR kinases.

Accordingly, the invention provides inter alia:

-   -   A method for the prophylaxis or treatment of a disease state or        condition mediated by a FGFR kinase, which method comprises        administering to a subject in need thereof a compound of the        formula (I) as defined herein.    -   A method for the prophylaxis or treatment of a disease state or        condition as described herein, which method comprises        administering to a subject in need thereof a compound of the        formula (I) as defined herein.    -   A method for the prophylaxis or treatment of cancer, which        method comprises administering to a subject in need thereof a        compound of the formula (I) as defined herein.    -   A method for alleviating or reducing the incidence of a disease        state or condition mediated by a FGFR kinase, which method        comprises administering to a subject in need thereof a compound        of the formula (I) as defined herein.    -   A method of inhibiting a FGFR kinase, which method comprises        contacting the kinase with a kinase-inhibiting compound of the        formula (I) as defined herein.    -   A method of modulating a cellular process (for example cell        division) by inhibiting the activity of a FGFR kinase using a        compound of the formula (I) as defined herein.    -   A compound of formula (I) as defined herein for use as a        modulator of a cellular process (for example cell division) by        inhibiting the activity of a FGFR kinase.    -   A compound of formula (I) as defined herein for use in the        prophylaxis or treatment of cancer, in particular the treatment        of cancer.    -   A compound of formula (I) as defined herein for use as a        modulator (e.g. inhibitor) of FGFR.    -   The use of a compound of formula (I) as defined herein for the        manufacture of a medicament for the prophylaxis or treatment of        a disease state or condition mediated by a FGFR kinase, the        compound having the formula (I) as defined herein.    -   The use of a compound of formula (I) as defined herein for the        manufacture of a medicament for the prophylaxis or treatment of        a disease state or condition as described herein.    -   The use of a compound of formula (I) as defined herein for the        manufacture of a medicament for the prophylaxis or treatment, in        particular the treatment, of cancer.    -   The use of a compound of formula (I) as defined herein for the        manufacture of a medicament for modulating (e.g. inhibiting) the        activity of FGFR.    -   Use of a compound of formula (I) as defined herein in the        manufacture of a medicament for modulating a cellular process        (for example cell division) by inhibiting the activity of a FGFR        kinase.    -   The use of a compound of the formula (I) as defined herein for        the manufacture of a medicament for prophylaxis or treatment of        a disease or condition characterised by up-regulation of a FGFR        kinase (e.g. FGFR1 or FGFR2 or FGFR3 or FGFR4).    -   The use of a compound of the formula (I) as defined herein for        the manufacture of a medicament for the prophylaxis or treatment        of a cancer, the cancer being one which is characterised by        up-regulation of a FGFR kinase (e.g. FGFR1 or FGFR2 or FGFR3 or        FGFR4).    -   The use of a compound of the formula (I) as defined herein for        the manufacture of a medicament for the prophylaxis or treatment        of cancer in a patient selected from a sub-population possessing        a genetic aberrations of FGFR3 kinase.    -   The use of a compound of the formula (I) as defined herein for        the manufacture of a medicament for the prophylaxis or treatment        of cancer in a patient who has been diagnosed as forming part of        a sub-population possessing a genetic aberrations of FGFR3        kinase.    -   A method for the prophylaxis or treatment of a disease or        condition characterised by up-regulation of a FGFR kinase (e.g.        FGFR1 or FGFR2 or FGFR3 or FGFR4), the method comprising        administering a compound of the formula (I) as defined herein.    -   A method for alleviating or reducing the incidence of a disease        or condition characterised by up-regulation of a FGFR kinase        (e.g. FGFR1 or FGFR2 or FGFR3 or FGFR4), the method comprising        administering a compound of the formula (I) as defined herein.    -   A method for the prophylaxis or treatment of (or alleviating or        reducing the incidence of) cancer in a patient suffering from or        suspected of suffering from cancer; which method comprises (i)        subjecting a patient to a diagnostic test to determine whether        the patient possesses a genetic aberrations of FGFR3 gene;        and (ii) where the patient does possess the said variant,        thereafter administering to the patient a compound of the        formula (I) as defined herein having FGFR3 kinase inhibiting        activity.    -   A method for the prophylaxis or treatment of (or alleviating or        reducing the incidence of) a disease state or condition        characterised by up-regulation of an FGFR kinase (e.g. FGFR1 or        FGFR2 or FGFR3 or FGFR4); which method comprises (i) subjecting        a patient to a diagnostic test to detect a marker characteristic        of up-regulation of a FGFR kinase (e.g. FGFR1 or FGFR2 or FGFR3        or FGFR4) and (ii) where the diagnostic test is indicative of        up-regulation of a FGFR kinase, thereafter administering to the        patient a compound of the formula (I) as defined herein having        FGFR kinase inhibiting activity.

In one embodiment, the disease mediated by FGFR kinases is a oncologyrelated disease (e.g. cancer). In one embodiment, the disease mediatedby FGFR kinases is a non-oncology related disease (e.g. any diseasedisclosed herein excluding cancer). In one embodiment the diseasemediated by FGFR kinases is a condition described herein. In oneembodiment the disease mediated by FGFR kinases is a skeletal conditiondescribed herein. Particular abnormalities in human skeletaldevelopment, include abnormal ossification of cranial sutures(craniosynostosis), Apert (AP) syndrome, Crouzon syndrome, Jackson-Weisssyndrome, Beare-Stevenson cutis gyrate syndrome, Pfeiffer syndrome,achondroplasia and thanatophoric dwarfism (also known as thanatophoricdysplasia).

Mutated Kinases

Drug resistant kinase mutations can arise in patient populations treatedwith kinase inhibitors.

These occur, in part, in the regions of the protein that bind to orinteract with the particular inhibitor used in therapy. Such mutationsreduce or increase the capacity of the inhibitor to bind to and inhibitthe kinase in question. This can occur at any of the amino acid residueswhich interact with the inhibitor or are important for supporting thebinding of said inhibitor to the target. An inhibitor that binds to atarget kinase without requiring the interaction with the mutated aminoacid residue will likely be unaffected by the mutation and will remainan effective inhibitor of the enzyme.

A study in gastric cancer patient samples showed the presence of twomutations in FGFR2, Ser167Pro in exon Ma and a splice site mutation940-2A-G in exon IIIc. These mutations are identical to the germlineactivating mutations that cause craniosynotosis syndromes and wereobserved in 13% of primary gastric cancer tissues studied. In additionactivating mutations in FGFR3 were observed in 5% of the patient samplestested and overexpression of FGFRs has been correlated with a poorprognosis in this patient group.

In addition there are chromosomal translocations or point mutations thathave been observed in FGFR which give rise to gain-of-function,over-expressed, or constitutively active biological states.

The compounds of the invention would therefore find particularapplication in relation to cancers which express a mutated moleculartarget such as FGFR. Diagnosis of tumours with such mutations could beperformed using techniques known to a person skilled in the art and asdescribed herein such as RTPCR and FISH.

It has been suggested that mutations of a conserved threonine residue atthe ATP binding site of FGFR would result in inhibitor resistance. Theamino acid valine 561 has been mutated to a methionine in FGFR1 whichcorresponds to previously reported mutations found in Abl (T315) andEGFR (T766) that have been shown to confer resistance to selectiveinhibitors. Assay data for FGFR1 V561M showed that this mutationconferred resistance to a tyrosine kinase inhibitor compared to that ofthe wild type.

Methods of Diagnosis

Prior to administration of a compound of the formula (I), a patient maybe screened to determine whether a disease or condition from which thepatient is or may be suffering is one which would be susceptible totreatment with a compound having activity against FGFR, and/or VEGFR.

For example, a biological sample taken from a patient may be analysed todetermine whether a condition or disease, such as cancer, that thepatient is or may be suffering from is one which is characterised by agenetic abnormality or abnormal protein expression which leads toup-regulation of the levels or activity of FGFR, and/or VEGFR or tosensitisation of a pathway to normal FGFR, and/or VEGFR activity, or toupregulation of these growth factor signalling pathways such as growthfactor ligand levels or growth factor ligand activity or to upregulationof a biochemical pathway downstream of FGFR, and/or VEGFR activation.

Examples of such abnormalities that result in activation orsensitisation of the FGFR, and/or VEGFR signal include loss of, orinhibition of apoptotic pathways, up-regulation of the receptors orligands, or presence of mutant variants of the receptors or ligands e.gPTK variants. Tumours with mutants of FGFR1, FGFR2 or FGFR3 or FGFR4 orup-regulation, in particular over-expression of FGFR1, orgain-of-function mutants of FGFR2 or FGFR3 may be particularly sensitiveto FGFR inhibitors.

For example, point mutations engendering gain-of-function in FGFR2 havebeen identified in a number of conditions. In particular activatingmutations in FGFR2 have been identified in 10% of endometrial tumours.

In addition, genetic aberrations of the FGFR3 receptor tyrosine kinasesuch as chromosomal translocations or point mutations resulting inectopically expressed or deregulated, constitutively active, FGFR3receptors have been identified and are linked to a subset of multiplemyelomas, bladder and cervical carcinomas. A particular mutation T6741of the PDGF receptor has been identified in imatinib-treated patients.In addition, a gene amplification of 8p12-p11.2 was demonstrated in ˜50%of lobular breast cancer (CLC) cases and this was shown to be linkedwith an increased expression of FGFR1. Preliminary studies with siRNAdirected against FGFR1, or a small molecule inhibitor of the receptor,showed cell lines harbouring this amplification to be particularlysensitive to inhibition of this signalling pathway.

Alternatively, a biological sample taken from a patient may be analysedfor loss of a negative regulator or suppressor of FGFR or VEGFR. In thepresent context, the term “loss” embraces the deletion of a geneencoding the regulator or suppressor, the truncation of the gene (forexample by mutation), the truncation of the transcribed product of thegene, or the inactivation of the transcribed product (e.g. by pointmutation) or sequestration by another gene product.

The term up-regulation includes elevated expression or over-expression,including gene amplification (i.e. multiple gene copies) and increasedexpression by a transcriptional effect, and hyperactivity andactivation, including activation by mutations. Thus, the patient may besubjected to a diagnostic test to detect a marker characteristic ofup-regulation of FGFR, and/or VEGFR. The term diagnosis includesscreening. By marker we include genetic markers including, for example,the measurement of DNA composition to identify mutations of FGFR, and/orVEGFR. The term marker also includes markers which are characteristic ofup regulation of FGFR and/or VEGFR, including enzyme activity, enzymelevels, enzyme state (e.g. phosphorylated or not) and mRNA levels of theaforementioned proteins.

The diagnostic tests and screens are typically conducted on a biologicalsample selected from tumour biopsy samples, blood samples (isolation andenrichment of shed tumour cells), stool biopsies, sputum, chromosomeanalysis, pleural fluid, peritoneal fluid, buccal spears, biopsy orurine.

Methods of identification and analysis of mutations and up-regulation ofproteins are known to a person skilled in the art. Screening methodscould include, but are not limited to, standard methods such asreverse-transcriptase polymerase chain reaction (RT-PCR) or in-situhybridization such as fluorescence in situ hybridization (FISH).

Identification of an individual carrying a mutation in FGFR, and/orVEGFR may mean that the patient would be particularly suitable fortreatment with a FGFR, and/or VEGFR inhibitor. Tumours maypreferentially be screened for presence of a FGFR, and/or VEGFR variantprior to treatment. The screening process will typically involve directsequencing, oligonucleotide microarray analysis, or a mutant specificantibody. In addition, diagnosis of tumours with such mutations could beperformed using techniques known to a person skilled in the art and asdescribed herein such as RT-PCR and FISH.

In addition, mutant forms of, for example FGFR or VEGFR2, can beidentified by direct sequencing of, for example, tumour biopsies usingPCR and methods to sequence PCR products directly as hereinbeforedescribed. The skilled artisan will recognize that all such well-knowntechniques for detection of the over expression, activation or mutationsof the aforementioned proteins could be applicable in the present case.

In screening by RT-PCR, the level of mRNA in the tumour is assessed bycreating a cDNA copy of the mRNA followed by amplification of the cDNAby PCR. Methods of PCR amplification, the selection of primers, andconditions for amplification, are known to a person skilled in the art.Nucleic acid manipulations and PCR are carried out by standard methods,as described for example in Ausubel, F. M. et al., eds. (2004) CurrentProtocols in Molecular Biology, John Wiley & Sons Inc., or Innis, M. A.et al., eds. (1990) PCR Protocols: a guide to methods and applications,Academic Press, San Diego. Reactions and manipulations involving nucleicacid techniques are also described in Sambrook et al., (2001), 3^(rd)Ed, Molecular Cloning: A Laboratory Manual, Cold Spring HarborLaboratory Press. Alternatively a commercially available kit for RT-PCR(for example Roche Molecular Biochemicals) may be used, or methodologyas set forth in U.S. Pat. Nos. 4,666,828; 4,683,202; 4,801,531;5,192,659, 5,272,057, 5,882,864, and 6,218,529 and incorporated hereinby reference. An example of an in-situ hybridisation technique forassessing mRNA expression would be fluorescence in-situ hybridisation(FISH) (see Angerer (1987) Meth. Enzymol., 152: 649).

Generally, in situ hybridization comprises the following major steps:(1) fixation of tissue to be analyzed; (2) prehybridization treatment ofthe sample to increase accessibility of target nucleic acid, and toreduce nonspecific binding; (3) hybridization of the mixture of nucleicacids to the nucleic acid in the biological structure or tissue; (4)post-hybridization washes to remove nucleic acid fragments not bound inthe hybridization, and (5) detection of the hybridized nucleic acidfragments. The probes used in such applications are typically labelled,for example, with radioisotopes or fluorescent reporters. Preferredprobes are sufficiently long, for example, from about 50, 100, or 200nucleotides to about 1000 or more nucleotides, to enable specifichybridization with the target nucleic acid(s) under stringentconditions. Standard methods for carrying out FISH are described inAusubel, F. M. et al., eds. (2004) Current Protocols in MolecularBiology, John Wiley & Sons Inc and Fluorescence In Situ Hybridization:Technical Overview by John M. S. Bartlett in Molecular Diagnosis ofCancer, Methods and Protocols, 2nd ed.; ISBN: 1-59259-760-2; March 2004,pps. 077-088; Series: Methods in Molecular Medicine.

Methods for gene expression profiling are described by (DePrimo et al.(2003), BMC Cancer, 3:3). Briefly, the protocol is as follows:double-stranded cDNA is synthesized from total RNA Using a (dT)24oligomer for priming first-strand cDNA synthesis, followed by secondstrand cDNA synthesis with random hexamer primers. The double-strandedcDNA is used as a template for in vitro transcription of cRNA usingbiotinylated ribonucleotides. cRNA is chemically fragmented according toprotocols described by Affymetrix (Santa Clara, Calif., USA), and thenhybridized overnight on Human Genome Arrays.

Alternatively, the protein products expressed from the mRNAs may beassayed by immunohistochemistry of tumour samples, solid phaseimmunoassay with microtitre plates, Western blotting, 2-dimensionalSDS-polyacrylamide gel electrophoresis, ELISA, flow cytometry and othermethods known in the art for detection of specific proteins. Detectionmethods would include the use of site specific antibodies. The skilledperson will recognize that all such well-known techniques for detectionof upregulation of FGFR, and/or VEGFR, or detection of FGFR, and/orVEGFR variants or mutants could be applicable in the present case.

Abnormal levels of proteins such as FGFR or VEGFR can be measured usingstandard enzyme assays, for example, those assays described herein.Activation or overexpression could also be detected in a tissue sample,for example, a tumour tissue. By measuring the tyrosine kinase activitywith an assay such as that from Chemicon International. The tyrosinekinase of interest would be immunoprecipitated from the sample lysateand its activity measured.

Alternative methods for the measurement of the over expression oractivation of FGFR or VEGFR including the isoforms thereof, include themeasurement of microvessel density. This can for example be measuredusing methods described by Orre and Rogers (Int J Cancer (1999), 84(2)101-8). Assay methods also include the use of markers, for example, inthe case of VEGFR these include CD31, CD34 and CD105.

Therefore all of these techniques could also be used to identify tumoursparticularly suitable for treatment with the compounds of the invention.

The compounds of the invention are particular useful in treatment of apatient having a mutated FGFR. The G697C mutation in FGFR3 is observedin 62% of oral squamous cell carcmonas and causes constitutiveactivation of the kinase activity. Activating mutations of FGFR3 havealso been identified in bladder carcinoma cases. These mutations were of6 kinds with varying degrees of prevelence: R248C, S249C, G372C, S373C,Y375C, K652Q. In addition, a Gly388Arg polymorphism in FGFR4 has beenfound to be associated with increased incidence and aggressiveness ofprostate, colon, lung, liver (HCC) and breast cancer. The compounds ofthe invention are particular useful in treatment of a patient having aFGFR3-TACC3 translocation.

Therefore in a further aspect the invention includes use of a compoundaccording to the invention for the manufacture of a medicament for thetreatment or prophylaxis of a disease state or condition in a patientwho has been screened and has been determined as suffering from, orbeing at risk of suffering from, a disease or condition which would besusceptible to treatment with a compound having activity against FGFR.

Particular mutations a patient is screened for include G697C, R248C,S249C, G372C, S373C, Y375C, K652Q mutations in FGFR3 and Gly388Argpolymorphism in FGFR4.

In another aspect the invention includes a compound of the invention foruse in the prophylaxis or treatment of cancer in a patient selected froma sub-population possessing a variant of the FGFR gene (for exampleG697C mutation in FGFR3 and Gly388Arg polymorphism in FGFR4).

MRI determination of vessel normalization (e.g. using MRI gradient echo,spin echo, and contrast enhancement to measure blood volume, relativevessel size, and vascular permeability) in combination with circulatingbiomarkers (circulating progenitor cells (CPCs), CECs, SDF1, and FGF2)may also be used to identify VEGFR2-resistant tumours for treatment witha compound of the invention.

Pharmaceutical Compositions and Combinations

In view of their useful pharmacological properties, the subjectcompounds may be formulated into various pharmaceutical forms foradministration purposes.

In one embodiment the pharmaceutical composition (e.g. formulation)comprises at least one active compound of the invention together withone or more pharmaceutically acceptable carriers, adjuvants, excipients,diluents, fillers, buffers, stabilisers, preservatives, lubricants, orother materials well known to those skilled in the art and optionallyother therapeutic or prophylactic agents.

To prepare the pharmaceutical compositions of this invention, aneffective amount of a compound of the present invention, as the activeingredient is combined in intimate admixture with a pharmaceuticallyacceptable carrier, which carrier may take a wide variety of formsdepending on the form of preparation desired for administration. Thepharmaceutical compositions can be in any form suitable for oral,parenteral, topical, intranasal, ophthalmic, otic, rectal,intra-vaginal, or transdermal administration. These pharmaceuticalcompositions are desirably in unitary dosage form suitable, preferably,for administration orally, rectally, percutaneously, or by parenteralinjection. For example, in preparing the compositions in oral dosageform, any of the usual pharmaceutical media may be employed, such as,for example, water, glycols, oils, alcohols and the like in the case oforal liquid preparations such as suspensions, syrups, elixirs andsolutions; or solid carriers such as starches, sugars, kaolin,lubricants, binders, disintegrating agents and the like in the case ofpowders, pills, capsules and tablets.

Because of their ease in administration, tablets and capsules representthe most advantageous oral dosage unit form, in which case solidpharmaceutical carriers are obviously employed. For parenteralcompositions, the carrier will usually comprise sterile water, at leastin large part, though other ingredients, to aid solubility for example,may be included. Injectable solutions, for example, may be prepared inwhich the carrier comprises saline solution, glucose solution or amixture of saline and glucose solution. Injectable suspensions may alsobe prepared in which case appropriate liquid carriers, suspending agentsand the like may be employed. In the compositions suitable forpercutaneous administration, the carrier optionally comprises apenetration enhancing agent and/or a suitable wetting agent, optionallycombined with suitable additives of any nature in minor proportions,which additives do not cause a significant deleterious effect to theskin. Said additives may facilitate the administration to the skinand/or may be helpful for preparing the desired compositions. Thesecompositions may be administered in various ways, e.g., as a transdermalpatch, as a spot-on, as an ointment. It is especially advantageous toformulate the aforementioned pharmaceutical compositions in dosage unitform for ease of administration and uniformity of dosage. Dosage unitform as used in the specification and claims herein refers to physicallydiscrete units suitable as unitary dosages, each unit containing apredetermined quantity of active ingredient calculated to produce thedesired therapeutic effect in association with the requiredpharmaceutical carrier. Examples of such dosage unit forms are tablets(including scored or coated tablets), capsules, pills, powder packets,wafers, injectable solutions or suspensions, teaspoonfuls,tablespoonfuls and the like, and segregated multiples thereof.

It is especially advantageous to formulate the aforementionedpharmaceutical compositions in dosage unit form for ease ofadministration and uniformity of dosage. Dosage unit form as used in thespecification and claims herein refers to physically discrete unitssuitable as unitary dosages, each unit containing a predeterminedquantity of active ingredient, calculated to produce the desiredtherapeutic effect, in association with the required pharmaceuticalcarrier.

Examples of such dosage unit forms are tablets (including scored orcoated tablets), capsules, pills, powder packets, wafers, injectablesolutions or suspensions, teaspoonfuls, tablespoonfuls and the like, andsegregated multiples thereof.

The compound of the invention is administered in an amount sufficient toexert its anti-tumour activity.

Those skilled in the art could easily determine the effective amountfrom the test results presented hereinafter. In general it iscontemplated that a therapeutically effective amount would be from 0.005mg/kg to 100 mg/kg body weight, and in particular from 0.005 mg/kg to 10mg/kg body weight. It may be appropriate to administer the required doseas single, two, three, four or more sub-doses at appropriate intervalsthroughout the day. Said sub-doses may be formulated as unit dosageforms, for example, containing 0.5 to 500 mg, in particular 1 mg to 500mg, more in particular 10 mg to 500 mg of active ingredient per unitdosage form.

Depending on the mode of administration, the pharmaceutical compositionwill preferably comprise from 0.05 to 99% by weight, more preferablyfrom 0.1 to 70% by weight, even more preferably from 0.1 to 50% byweight of the compound of the present invention, and, from 1 to 99.95%by weight, more preferably from 30 to 99.9% by weight, even morepreferably from 50 to 99.9% by weight of a pharmaceutically acceptablecarrier, all percentages being based on the total weight of thecomposition.

As another aspect of the present invention, a combination of a compoundof the present invention with another anticancer agent is envisaged,especially for use as a medicine, more specifically for use in thetreatment of cancer or related diseases.

For the treatment of the above conditions, the compounds of theinvention may be advantageously employed in combination with one or moreother medicinal agents, more particularly, with other anti-cancer agentsor adjuvants in cancer therapy. Examples of anti-cancer agents oradjuvants (supporting agents in the therapy) include but are not limitedto:

-   -   platinum coordination compounds for example cisplatin optionally        combined with amifostine, carboplatin or oxaliplatin;    -   taxane compounds for example paclitaxel, paclitaxel protein        bound particles (Abraxane™) or docetaxel;    -   topoisomerase I inhibitors such as camptothecin compounds for        example irinotecan, SN-38, topotecan, topotecan hcl;    -   topoisomerase II inhibitors such as anti-tumour        epipodophyllotoxins or podophyllotoxin derivatives for example        etoposide, etoposide phosphate or teniposide;    -   anti-tumour vinca alkaloids for example vinblastine, vincristine        or vinorelbine;    -   anti-tumour nucleoside derivatives for example 5-fluorouracil,        leucovorin, gemcitabine, gemcitabine hcl, capecitabine,        cladribine, fludarabine, nelarabine;    -   alkylating agents such as nitrogen mustard or nitrosourea for        example cyclophosphamide, chlorambucil, carmustine, thiotepa,        mephalan (melphalan), lomustine, altretamine, busulfan,        dacarbazine, estramustine, ifosfamide optionally in combination        with mesna, pipobroman, procarbazine, streptozocin,        telozolomide, uracil;    -   anti-tumour anthracycline derivatives for example daunorubicin,        doxorubicin optionally in combination with dexrazoxane, doxil,        idarubicin, mitoxantrone, epirubicin, epirubicin hcl,        valrubicin;    -   molecules that target the IGF-1 receptor for example        picropodophilin;    -   tetracarcin derivatives for example tetrocarcin A;    -   glucocorticoids for example prednisone;    -   antibodies for example trastuzumab (HER2 antibody), rituximab        (CD20 antibody), gemtuzumab, gemtuzumab ozogamicin, cetuximab,        pertuzumab, bevacizumab, alemtuzumab, eculizumab, ibritumomab        tiuxetan, nofetumomab, panitumumab, tositumomab, CNTO 328;    -   estrogen receptor antagonists or selective estrogen receptor        modulators or inhibitors of estrogen synthesis for example        tamoxifen, fulvestrant, toremifene, droloxifene, faslodex,        raloxifene or letrozole;    -   aromatase inhibitors such as exemestane, anastrozole, letrazole,        testolactone and vorozole;    -   differentiating agents such as retinoids, vitamin D or retinoic        acid and retinoic acid metabolism blocking agents (RAMBA) for        example accutane;    -   DNA methyl transferase inhibitors for example azacytidine or        decitabine;    -   antifolates for example premetrexed disodium;    -   antibiotics for example antinomycin D, bleomycin, mitomycin C,        dactinomycin, carminomycin, daunomycin, levamisole, plicamycin,        mithramycin;    -   antimetabolites for example clofarabine, aminopterin, cytosine        arabinoside or methotrexate, azacitidine, cytarabine,        floxuridine, pentostatin, thioguanine;    -   apoptosis inducing agents and antiangiogenic agents such as        Bcl-2 inhibitors for example YC137, BH 312, ABT 737, gossypol,        HA 14-1, TW 37 or decanoic acid;    -   tubuline-binding agents for example combrestatin, colchicines or        nocodazole;    -   kinase inhibitors (e.g. EGFR (epithelial growth factor receptor)        inhibitors, MTKI (multi target kinase inhibitors), mTOR        inhibitors, cmet inhibitors) for example flavoperidol, imatinib        mesylate, erlotinib, gefitinib, dasatinib, lapatinib, lapatinib        ditosylate, sorafenib, sunitinib, sunitinib maleate,        temsirolimus,        6-{difluoro[6-(1-methyl-1H-pyrazol-4-yl)[1,2,4]triazolo[4,3-b]pyridazin-3-yl]methyl}quinoline        or a pharmaceutically ac ceptable salt thereof,        6-[difluoro(6-pyridin-4-yl[1,2,4]triazolo[4,3-b]pyridazin-3-yl)methyl]quinoline        or a pharmaceutically acceptable salt thereof;    -   farnesyltransferase inhibitors for example tipifarnib;    -   histone deacetylase (HDAC) inhibitors for example sodium        butyrate, suberoylanilide hydroxamide acid (SAHA), depsipeptide        (FR 901228), NVP-LAQ824, R306465, JNJ-26481585, trichostatin A,        vorinostat;    -   Inhibitors of the ubiquitin-proteasome pathway for example        PS-341, MLN 0.41 or bortezomib;    -   Yondelis;    -   Telomerase inhibitors for example telomestatin;    -   Matrix metalloproteinase inhibitors for example batimastat,        marimastat, prinostat or metastat.    -   Recombinant interleukins for example aldesleukin, denileukin        diftitox, interferon alfa 2a, interferon alfa 2b, peginterferon        alfa 2b    -   MAPK inhibitors    -   Retinoids for example alitretinoin, bexarotene, tretinoin    -   Arsenic trioxide    -   Asparaginase    -   Steroids for example dromostanolone propionate, megestrol        acetate, nandrolone (decanoate, phenpropionate), dexamethasone    -   Gonadotropin releasing hormone agonists or antagonists for        example abarelix, goserelin acetate, histrelin acetate,        leuprolide acetate    -   Thalidomide, lenalidomide    -   Mercaptopurine, mitotane, pamidronate, pegademase, pegaspargase,        rasburicase    -   BH3 mimetics for example ABT-737    -   MEK inhibitors for example PD98059, AZD6244, CI-1040    -   colony-stimulating factor analogs for example filgrastim,        pegfilgrastim, sargramostim; erythropoietin or analogues thereof        (e.g. darbepoetin alfa); interleukin 11; oprelvekin;        zoledronate, zoledronic acid; fentanyl; bisphosphonate;        palifermin.    -   a steroidal cytochrome P450 17alpha-hydroxylase-17,20-lyase        inhibitor (CYP17), e.g. abiraterone, abiraterone acetate.

In one embodiment, the present invention relates to a combination of acompound of formula (I), a pharmaceutically acceptable salt thereof or asolvate thereof, or any sub-groups and examples thereof, and6-{difluoro[6-(1-methyl-1H-pyrazol-4-yl)[1,2,4]triazolo[4,3-b]pyridazin-3-yl]methyl}quinolineor a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to a combination of acompound of formula (I), a pharmaceutically acceptable salt thereof or asolvate thereof, or any sub-groups and examples thereof, and6-[difluoro(6-pyridin-4-yl[1,2,4]triazolo[4,3-b]pyridazin-3-yl)methyl]quinolineor a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to a pharmaceuticalcomposition comprising a compound of formula (I), a pharmaceuticallyacceptable salt thereof or a solvate thereof, or any sub-groups andexamples thereof, and6-{difluoro[6-(1-methyl-1H-pyrazol-4-yl)[1,2,4]triazolo[4,3-b]pyridazin-3-yl]methyl}quinolineor a pharmaceutically acceptable salt thereof.

In one embodiment, the present invention relates to a pharmaceuticalcomposition comprising a compound of formula (I), a pharmaceuticallyacceptable salt thereof or a solvate thereof, or any sub-groups andexamples thereof, and6-[difluoro(6-pyridin-4-yl[1,2,4]triazolo[4,3-b]pyridazin-3-yl)methyl]quinolineor a pharmaceutically acceptable salt thereof.

The compounds of the present invention also have therapeuticapplications in sensitising tumour cells for radiotherapy andchemotherapy.

Hence the compounds of the present invention can be used as“radiosensitizer” and/or “chemosensitizer” or can be given incombination with another “radiosensitizer” and/or “chemosensitizer”.

The term “radiosensitizer”, as used herein, is defined as a molecule,preferably a low molecular weight molecule, administered to animals intherapeutically effective amounts to increase the sensitivity of thecells to ionizing radiation and/or to promote the treatment of diseaseswhich are treatable with ionizing radiation.

The term “chemosensitizer”, as used herein, is defined as a molecule,preferably a low molecular weight molecule, administered to animals intherapeutically effective amounts to increase the sensitivity of cellsto chemotherapy and/or promote the treatment of diseases which aretreatable with chemotherapeutics.

Several mechanisms for the mode of action of radiosensitizers have beensuggested in the literature including: hypoxic cell radiosensitizers(e.g., 2-nitroimidazole compounds, and benzotriazine dioxide compounds)mimicking oxygen or alternatively behave like bioreductive agents underhypoxia; non-hypoxic cell radiosensitizers (e.g., halogenatedpyrimidines) can be analogoues of DNA bases and preferentiallyincorporate into the DNA of cancer cells and thereby promote theradiation-induced breaking of DNA molecules and/or prevent the normalDNA repair mechanisms; and various other potential mechanisms of actionhave been hypothesized for radiosensitizers in the treatment of disease.

Many cancer treatment protocols currently employ radiosensitizers inconjunction with radiation of x-rays. Examples of x-ray activatedradiosensitizers include, but are not limited to, the following:metronidazole, misonidazole, desmethylmisonidazole, pimonidazole,etanidazole, nimorazole, mitomycin C, RSU 1069, SR 4233, E09, RB 6145,nicotinamide, 5-bromodeoxyuridine (BUdR), 5-iododeoxyuridine (IUdR),bromodeoxycytidine, fluorodeoxyuridine (FudR), hydroxyurea, cisplatin,and therapeutically effective analogs and derivatives of the same.

Photodynamic therapy (PDT) of cancers employs visible light as theradiation activator of the sensitizing agent. Examples of photodynamicradiosensitizers include the following, but are not limited to:hematoporphyrin derivatives, Photofrin, benzoporphyrin derivatives, tinetioporphyrin, pheoborbide-a, bacteriochlorophyll-a, naphthalocyanines,phthalocyanines, zinc phthalocyanine, and therapeutically effectiveanalogs and derivatives of the same. Radiosensitizers may beadministered in conjunction with a therapeutically effective amount ofone or more other compounds, including but not limited to: compoundswhich promote the incorporation of radiosensitizers to the target cells;compounds which control the flow of therapeutics, nutrients, and/oroxygen to the target cells; chemotherapeutic agents which act on thetumour with or without additional radiation; or other therapeuticallyeffective compounds for treating cancer or other diseases.

Chemosensitizers may be administered in conjunction with atherapeutically effective amount of one or more other compounds,including but not limited to: compounds which promote the incorporationof chemosensitizers to the target cells; compounds which control theflow of therapeutics, nutrients, and/or oxygen to the target cells;chemotherapeutic agents which act on the tumour or other therapeuticallyeffective compounds for treating cancer or other disease. Calciumantagonists, for example verapamil, are found useful in combination withantineoplastic agents to establish chemosensitivity in tumor cellsresistant to accepted chemotherapeutic agents and to potentiate theefficacy of such compounds in drug-sensitive malignancies.

In view of their useful pharmacological properties, the components ofthe combinations according to the invention, i.e. the one or more othermedicinal agent and the compound according to the present invention maybe formulated into various pharmaceutical forms for administrationpurposes. The components may be formulated separately in individualpharmaceutical compositions or in a unitary pharmaceutical compositioncontaining all components.

The present invention therefore also relates to a pharmaceuticalcomposition comprising the one or more other medicinal agent and thecompound according to the present invention together with apharmaceutical carrier.

The present invention further relates to the use of a combinationaccording to the invention in the manufacture of a pharmaceuticalcomposition for inhibiting the growth of tumour cells.

The present invention further relates to a product containing as firstactive ingredient a compound according to the invention and as furtheractive ingredient one or more anticancer agent, as a combinedpreparation for simultaneous, separate or sequential use in thetreatment of patients suffering from cancer.

The one or more other medicinal agents and the compound according to thepresent invention may be administered simultaneously (e.g. in separateor unitary compositions) or sequentially in either order. In the lattercase, the two or more compounds will be administered within a period andin an amount and manner that is sufficient to ensure that anadvantageous or synergistic effect is achieved. It will be appreciatedthat the preferred method and order of administration and the respectivedosage amounts and regimes for each component of the combination willdepend on the particular other medicinal agent and compound of thepresent invention being administered, their route of administration, theparticular tumour being treated and the particular host being treated.The optimum method and order of administration and the dosage amountsand regime can be readily determined by those skilled in the art usingconventional methods and in view of the information set out herein.

The weight ratio of the compound according to the present invention andthe one or more other anticancer agent(s) when given as a combinationmay be determined by the person skilled in the art. Said ratio and theexact dosage and frequency of administration depends on the particularcompound according to the invention and the other anticancer agent(s)used, the particular condition being treated, the severity of thecondition being treated, the age, weight, gender, diet, time ofadministration and general physical condition of the particular patient,the mode of administration as well as other medication the individualmay be taking, as is well known to those skilled in the art.Furthermore, it is evident that the effective daily amount may belowered or increased depending on the response of the treated subjectand/or depending on the evaluation of the physician prescribing thecompounds of the instant invention. A particular weight ratio for thepresent compound of formula (I) and another anticancer agent may rangefrom 1/10 to 10/1, more in particular from 1/5 to 5/1, even more inparticular from 1/3 to 3/1.

The platinum coordination compound is advantageously administered in adosage of 1 to 500 mg per square meter (mg/m²) of body surface area, forexample 50 to 400 mg/m², particularly for cisplatin in a dosage of about75 mg/m² and for carboplatin in about 300 mg/m² per course of treatment.

The taxane compound is advantageously administered in a dosage of 50 to400 mg per square meter (mg/m²) of body surface area, for example 75 to250 mg/m², particularly for paclitaxel in a dosage of about 175 to 250mg/m² and for docetaxel in about 75 to 150 mg/m² per course oftreatment.

The camptothecin compound is advantageously administered in a dosage of0.1 to 400 mg per square meter (mg/m²) of body surface area, for example1 to 300 mg/m², particularly for irinotecan in a dosage of about 100 to350 mg/m² and for topotecan in about 1 to 2 mg/m² per course oftreatment.

The anti-tumour podophyllotoxin derivative is advantageouslyadministered in a dosage of 30 to 300 mg per square meter (mg/m²) ofbody surface area, for example 50 to 250 mg/m², particularly foretoposide in a dosage of about 35 to 100 mg/m² and for teniposide inabout 50 to 250 mg/m² per course of treatment.

The anti-tumour vinca alkaloid is advantageously administered in adosage of 2 to 30 mg per square meter (mg/m²) of body surface area,particularly for vinblastine in a dosage of about 3 to 12 mg/m², forvincristine in a dosage of about 1 to 2 mg/m², and for vinorelbine indosage of about 10 to 30 mg/m² per course of treatment.

The anti-tumour nucleoside derivative is advantageously administered ina dosage of 200 to 2500 mg per square meter (mg/m²) of body surfacearea, for example 700 to 1500 mg/m², particularly for 5-FU in a dosageof 200 to 500 mg/m², for gemcitabine in a dosage of about 800 to 1200mg/m² and for capecitabine in about 1000 to 2500 mg/m² per course oftreatment.

The alkylating agents such as nitrogen mustard or nitrosourea isadvantageously administered in a dosage of 100 to 500 mg per squaremeter (mg/m²) of body surface area, for example 120 to 200 mg/m²,particularly for cyclophosphamide in a dosage of about 100 to 500 mg/m²,for chlorambucil in a dosage of about 0.1 to 0.2 mg/kg, for carmustinein a dosage of about 150 to 200 mg/m², and for lomustine in a dosage ofabout 100 to 150 mg/m² per course of treatment.

The anti-tumour anthracycline derivative is advantageously administeredin a dosage of 10 to 75 mg per square meter (mg/m²) of body surfacearea, for example 15 to 60 mg/m², particularly for doxorubicin in adosage of about 40 to 75 mg/m², for daunorubicin in a dosage of about 25to 45 mg/m², and for idarubicin in a dosage of about 10 to 15 mg/m² percourse of treatment.

The antiestrogen agent is advantageously administered in a dosage ofabout 1 to 100 mg daily depending on the particular agent and thecondition being treated. Tamoxifen is advantageously administered orallyin a dosage of 5 to 50 mg, preferably 10 to 20 mg twice a day,continuing the therapy for sufficient time to achieve and maintain atherapeutic effect. Toremifene is advantageously administered orally ina dosage of about 60 mg once a day, continuing the therapy forsufficient time to achieve and maintain a therapeutic effect.Anastrozole is advantageously administered orally in a dosage of about 1mg once a day. Droloxifene is advantageously administered orally in adosage of about 20-100 mg once a day. Raloxifene is advantageouslyadministered orally in a dosage of about 60 mg once a day. Exemestane isadvantageously administered orally in a dosage of about 25 mg once aday.

Antibodies are advantageously administered in a dosage of about 1 to 5mg per square meter (mg/m²) of body surface area, or as known in theart, if different. Trastuzumab is advantageously administered in adosage of 1 to 5 mg per square meter (mg/m²) of body surface area,particularly 2 to 4 mg/m² per course of treatment.

These dosages may be administered for example once, twice or more percourse of treatment, which may be repeated for example every 7, 14, 21or 28 days.

The compounds of formula (I), the pharmaceutically acceptable additionsalts, in particular pharmaceutically acceptable acid addition salts,and stereoisomeric forms thereof can have valuable diagnostic propertiesin that they can be used for detecting or identifying the formation of acomplex between a labelled compound and other molecules, peptides,proteins, enzymes or receptors.

The detecting or identifying methods can use compounds that are labelledwith labelling agents such as radioisotopes, enzymes, fluorescentsubstances, luminous substances, etc. Examples of the radioisotopesinclude ¹²⁵I, ¹³¹I, ¹³¹H and ¹⁴C. Enzymes are usually made detectable byconjugation of an appropriate substrate which, in turn catalyses adetectable reaction. Examples thereof include, for example,beta-galactosidase, beta-glucosidase, alkaline phosphatase, peroxidaseand malate dehydrogenase, preferably horseradish peroxidase. Theluminous substances include, for example, luminol, luminol derivatives,luciferin, aequorin and luciferase. Biological samples can be defined asbody tissue or body fluids. Examples of body fluids are cerebrospinalfluid, blood, plasma, serum, urine, sputum, saliva and the like.

General Synthetic Routes

The following examples illustrate the present invention but are examplesonly and are not intended to limit the scope of the claims in any way.

Experimental Part

Hereinafter, the term ‘DCM’ means dichloromethane, ‘Me’ means methyl,‘Et’ means ethyl, ‘MeOH’ means methanol, ‘DMF’ means dimethylformamide,‘Et₂O’ means diethyl ether, ‘EtOAc’ means ethyl acetate, ‘ACN’ meansacetonitrile, ‘H₂O’ means water, ‘THF’ means tetrahydrofuran, ‘MgSO₄’means magnesium sulfate, ‘NH₄OH’ means ammoniumhydroxide, ‘K₂CO₃’ meansdipotassium carbonate, ‘MgCl₂’ means magnesium chloride, ‘iPrNH₂’ meansisopropylamine, ‘NH₄HCO₃’ means ammonium bicarbonate, ‘DMSO’ meansdimethyl sulfoxide, ‘EDTA’ means ethylenediaminetetraacetic acid, ‘NADP’means nicotinamide adenine dinucleotide phosphate, ‘SFC’ meanssupercritical fluid chromatography, ‘MP’ means melting point.

A. Preparation of the Intermediates

Intermediate 1 orN-(3,5-dimethoxyphenyl)-N′-(1-methylethyl)-N-[3-(1-methyl-1H-pyrazol-4-yl)quinoxalin-6-yl]ethane-1,2-diamineis described as compound 4 in WO2011/135376 and can be preparedaccording to the protocols described therein for compound 4.

Intermediate 2 orN-(3,5-dimethoxyphenyl)-N′-(1-methylethyl)-N-[3-(1H-pyrazol-4-yl)quinoxalin-6-yl]ethane-1,2-diamineis described as free base compound 137 in WO2011/135376 and can beprepared according to the protocols described therein for compound 137.

Intermediate 3 orN-(3,5-dimethoxyphenyl)-N′-(1-methyl)-N-[3-(1-ethyl-1H-pyrazol-4-yl)quinoxalin-6-yl]ethane-1,2-diamineis described as compound 449 in WO2011/135376 and can be preparedaccording to the protocols described therein for compound 449.

Intermediate 4 orN-(3,5-dimethoxyphenyl)-N′-(1-methylethyl)-N-[3-(1-ethyl-1H-pyrazol-4-yl)quinoxalin-6-yl]ethane-1,2-diamineis described as free base or HCl salt as compound 135 in WO2011/135376and can be prepared according to the protocols described therein forcompound 135.

Intermediate 5 orN-(3,5-dimethoxyphenyl)-N′-(1-methylethyl)-N-[3-(1-methyl-1H-pyrazol-4-yl)quinoxalin-6-yl]propane-1,3-diamineis described as compound 382 in WO2011/135376 and can be preparedaccording to the protocols described therein for compound 382.

Intermediate 6 or 7-bromo-2-(1-methyl-1H-pyrazol-4-yl)-quinoxaline isdescribed as intermediate 2 in WO2011/135376 and can be preparedaccording to the protocols described therein for intermediate 2.

WO2011/135376 is incorporated herein by reference.

EXAMPLE A1

a) Preparation of Intermediate 7

A mixture of intermediate 6 (5 g; 17 mmol),2-fluoro-3,5-dimethoxyaniline (3.6 g; 21 mmol), sodium tert-butoxide (5g; 52 mmol) and rac-bis(diphenylphosphino)-1,1′-binaphthyl (0.54 g; 0.87mmol) in dioxane (100 mL) was degassed at room temperature undernitrogen flow. After 10 minutes, palladium (II) acetate (388 mg; 1.7mmol) was added portionwise at room temperature under nitrogen flow. Thereaction mixture was heated at 95° C. for 5 hours. The reaction mixturewas cooled to room temperature and poured onto iced water and DCM. Themixture was filtered through a pad of Celite®. The organic layer wasseparated, dried over MgSO₄, filtered and evaporated to dryness. Theresidue was crystallized from diethylether and the precipitate wasfiltered off, dried under vacuum to give 4 g (61%) of intermediate 7.

b) Preparation of Intermediate 8

Sodium hydride (0.21 g; 5.35 mmol) was added to a solution ofintermediate 7 (0.7 g; 1.85 mmol) in DMF (25 mL) at 5° C. under nitrogenflow. The mixture was stirred at 5° C. for 1 hour.(2-Bromoethoxy)-tert-butyldimethylsilane (0.51 mL; 2.40 mmol) was addeddropwise at 5° C. under nitrogen flow and the reaction mixture wasstirred at room temperature for 24 hours. The mixture was poured intocooled water and the product was extracted with EtOAc. The organic layerwas washed with H₂O, dried over MgSO₄, filtered and evaporated to give1.2 g (quant.) of intermediate 8. The crude product was used without anypurification in the next step.

c) Preparation of Intermediate 9

Tetrabutyl ammonium fluoride (1M in THF) (2 mL; 2 mmol) was added to asolution of intermediate 8 (1 g; 1.85 mmol) in THF (20 mL) and thereaction mixture was stirred for 3 hours at room temperature. Thereaction mixture was partitioned between water and EtOAc. The organiclayer was washed with brine, dried over MgSO₄, filtered and evaporatedto dryness. The residue (1.2 g) was purified by chromatography oversilica gel (irregular SiOH, 15-40 μm; 80 g; eluent: 98% DCM, 2% MeOH,0.1% NH₄OH). The pure fractions were collected and the solvent wasevaporated. The residue (500 mg) was crystallized from diethylether. Theprecipitate was filtered and dried to give 410 mg (52%) of intermediate9. MP: 172° C. (K).

d) Preparation of Intermediate 10

Methanesulfonyl chloride (0.3 mL; 3.88 mmol) was added dropwise at 5° C.to a solution of intermediate 9 (547 mg; 1.29 mmol) and triethylamine(0.9 mL; 6.46 mmol) in DCM (15 mL). The reaction mixture was stirred atthis temperature for 1 hour, diluted with DCM and poured onto 10%aqueous solution of K₂CO₃. The organic layer was decanted, dried overMgSO₄, filtered and evaporated to give 850 mg (>100%) of intermediate10. The crude product was used without purification in the next step.

e) Preparation of Intermediate 11

A mixture of intermediate 10 (0.648 g; 1.29 mmol) and isopropylamine(2.4 mL; 28 mmol) in CH₃CN (15 mL) was heated at 100° C. for 24 hours ina sealed tube. The reaction mixture was cooled to room temperature,diluted with DCM and poured onto water. The organic layer was decanted,dried over MgSO₄, filtered and evaporated to dryness. The residue waspurified by chromatography over silica gel (irregular SiOH; 24 g;gradient: from 3% MeOH, 97% DCM to 10% MeOH, 90% DCM). The purefractions were collected and evaporated to give 452 mg (75%) ofintermediate 11.

B. Preparation of the Compounds of Formula (I)

EXAMPLE B1

Preparation of Compound 1

A solution ofN-(3,5-dimethoxyphenyl)-N′-(1-methylethyl)-N-[3-(1-methyl-1H-pyrazol-4-yl)quinoxalin-6-yl]ethane-1,2-diamine(intermediate 1) (0.42 g; 0.9 mmol), formaldehyde (37% solution inwater; 0.21 mL; 2.8 mmol) in dioxane (8 mL) was stirred at roomtemperature for 3 days. Water and EtOAc were added. The organic layerwas decanted, washed with water, dried over MgSO₄ and evaporated todryness. The residue (0.52 g) was purified by chromatography over silicagel (Stationary phase: Spherical bare silica 5 μm 150×30.0 mm, Mobilephase: Gradient from 0.2% NH₄OH, 98% DCM, 2% MeOH to 1.2% NH₄OH, 88%DCM, 12% MeOH). The fractions containing the desired product werecollected and evaporated to dryness. The residue (0.37 g) wascrystallized from a mixture of MeOH and Et₂O. The precipitate wasfiltered off and dried, yielding 0.27 g (64%) of compound 1 (MP: 190° C.(DSC)).

EXAMPLE B2

Preparation of Compound 2

A solution ofN-(3,5-dimethoxyphenyl)-N′-(1-methylethyl)-N-[3-(1H-pyrazol-4-yl)quinoxalin-6-yl]ethane-1,2-diamine(intermediate 2) (0.24 g; 0.52 mmol), formaldehyde (37% solution inwater; 0.12 mL; 1.55 mmol) in dioxane (8 mL) was stirred at roomtemperature for 3 days without transformation. K₂CO₃ (0.22 g; 1.55 mmol)was added and the solution was further stirred at room temperature for 3days. The organic layer was extracted, dried over MgSO₄ and evaporatedto dryness. The residue (0.176 g) was purified by chromatography oversilica gel (Stationary phase: Spherical bare silica 5 μm 150×30.0 mm,Mobile phase: Gradient from 0.2% NH₄OH, 98% DCM, 2% MeOH to 1.3% NH₄OH,87% DCM, 13% MeOH). The fractions containing the desired product werecollected and evaporated to dryness. The residue (79 mg) wasfreeze-dried with acetonitrile/water 20/80 to give 66 mg (34%) ofcompound 2 as a yellow gummy powder.

EXAMPLE B3

Preparation of Compound 3 and 4

50 μM of intermediate 1 was incubated at 37° C. with the 12,000 gfraction from rat liver for 60 minutes at 1 mg/ml protein. A stocksolution of 10 mM of intermediate 1 in methanol was prepared and thiswas diluted 200-fold (0.25 ml in 50 ml) in the incubation medium (finalmethanol concentration 0.5% in the incubate). The incubation buffercontained 1 mM EDTA, 5 mM MgCl₂, and 100 mM potassium phosphate buffer(pH 7.4). Reaction was initiated by addition of NADP (1 mM finalconcentration). Incubation was stopped by flash freezing on dry ice.

The resulting metabolites were initially extracted using ethyl acetate.The metabolite fraction was evaporated to dryness, reconstituted inDMSO: water (1:1, v/v) and separated using reverse phase UPLC.Separation was achieved using two Interchim Strategy C18-2, 2.2 μm, (150mm×3.0 mm ID) columns using solvent A with a linear gradient of 5-70% Bover 20 minutes at 0.8 ml/min. Solvents consisted of Solvent A, 25 mMAmmonium acetate pH4.0 and Solvent B, Acetonitrile/methanol (60/40,v/v). Peak fractions corresponding to the desired products werecollected, and evaporated to dryness, yielding compound 3 and 4.

Compound 3 can also be prepared according to the protocol described inExample 1 starting from compound 645 of WO2011/135376.

Alternatively, compound 3 and 4 were also prepared as follows:

Boron tribromide (1M in DCM; 6 mL; 6 mmol) was added dropwise to asolution of compound 1 (485 mg; 1.06 mmol) in DCM (25 mL) at 5° C. undernitrogen flow. The solution was allowed raise to room temperature slowlyand was stirred for 1.5 hour. The reaction mixture was diluted with DCM,poured onto brine and basified with solid K₂CO₃. The organic layer wasseparated, washed with brine, dried over MgSO₄, filtered and evaporatedto dryness. The residue was purified by chromatography over silica gel(irregular SiOH, 40 g; mobile phase: gradient from 0.5% NH₄OH, 94.5%DCM, 5% MeOH to 0.5% NH₄OH, 89.5 DCM, 10% MeOH). The fractionscontaining the product were collected and evaporated to dryness yielding110 mg (23%) of compound 1 and 287 mg of a mixture of compounds 3 and 4.This latter fraction was purified by achiral SFC (Chiralpak AD-H 5 μm250*30 mn; mobile phase: 0.3% isopropylamine, 70% CO₂, 30% MeOH). Thepure fractions were collected, concentrated and crystallized fromEt₂O/ACN. The precipitates were filtered to afford 53 mg (11%) ofcompound 3 (MP: 255° C., K) and 148 mg (31%) of compound 4 (MP: 256° C.,K).

EXAMPLE B4

A stock solution of 2 mM of intermediate 1 was prepared in methanol andthis was diluted 200-fold in the incubation medium (final methanolconcentration 0.5% in the incubate). Incubation was done at 37° C. withthe 12,000 g fraction from rat liver for 60 minutes at 1 mg/ml protein.The incubation buffer contained 1 mM EDTA, 5 mM MgCl₂, and 100 mMpotassium phosphate buffer (pH 7.4). Reaction was initiated by additionof NADP (1 mM final concentration). Incubation was stopped by flashfreezing on dry ice.

The resulting incubation (1 ml) was mixed with 5 volumes ofacetonitrile, vortex mixed and sonicated for 10 minutes. Protein wasremoved by centrifugation at 3200 rpm at 8° C. for 30 minutes. Thesupernatant was removed and evaporated to dryness under a stream ofnitrogen at 30° C. The extract was reconstituted in acetonitrile/water(1:1, v/v). The sample was analysed as follows:

UPLC with MS Detection

-   -   Ultra performance liquid chromatography Pump        -   Acquity Binary Solvent Manager/Waters 2777 CTC-Pal-injector    -   UV detector:        -   Waters Acquity PDA    -   MS Detector:        -   Waters G2(S) QToF MS/Thermo LTQ-Orbitrap    -   Data System:        -   Waters Masslynx 4.1

Operating Conditions:

-   -   Column:        -   Interchim, Strategy C18-2, 2.2 μm 2×(150 mm×3.0 mm ID)    -   Column temperature:        -   T=60° C.    -   Sample temperature:        -   T=10° C.    -   Mobile phase:        -   Solvent A: 0.025 M ammonium acetate pH 4.0        -   Solvent B: 60/40 (v/v) acetonitrile/methanol    -   Elution mode:        -   linear gradient:

Time (min) 0 5 22 22.5 25 25.5 % A 95 80 50 0 0 95 % B 5 20 50 100 100 5

-   -   Run Time: 30 min    -   Flow: 0.8 ml/min

Syringe Infusion:

-   -   Mobile phase: Acetonitrile:water (1:1, v/v)    -   Flow: 5 μl/min        Detection Conditions        MS Conditions—Waters Synapt g2 and g2s Mass Spectrometer    -   MS analysis was carried out using Waters SYNAPT G2 and G2S mass        spectrometers, equipped with a dual electrospray ionisation        probe and was operated in high resolution, positive ion mode.        The capillary voltage was set at 3 kV and the cone at 40V. The        source temperature was 120° C., desolvation temperature 400° C.        The mass spectrometer was calibrated with a Sodium Formate        solution delivered through the Sample Spray. The LockSpray™ ESI        probe provided an independent source of the lock mass calibrant        Leucine Enkephaline. The Leucine ion at m/z 556.2771 was used as        lock mass in full MS as well as in MSMS-mode. The QTOF data (MS,        MSMS) were acquired in the centroid mode with a variable scan        time (0.5-1.0 sec). All data were processed using Masslynx        software.        MS Conditions—Thermo Ltq-Orbitrap Mass Spectrometer    -   The LTQ-Orbitrap mass spectrometer was equipped with an        electrospray ionisation source operated in the positive ion        mode. Accurate mass measurements were obtained using external        calibration or a lock mass calibration (lock mass ion at m/z        391.2843). The source parameters were tuned for maximum        sensitivity using a 10 ng/μL unchanged drug standard solution.        The same solution was used to define the optimal collision        energy employed during MS' fragmentation. Metabolites were        selected for MS' fragmentation out of the LC-MS trace using data        dependent scanning. Data were acquired in the centroid mode and        processed using XCalibur software.

In the above experiment compound 4 ([MH]+m/z 445), compound 3 ([MH]+m/z445) and compound 5 ([MH]+m/z 431) were detected.

EXAMPLE B5

Preparation of Compound 6

A solution of intermediate 3 (292 mg; 0.675 mmol), formaldehyde (37%solution in water; 151 μL; 2.02 mmol) in 1,4-dioxane (5.48 mL) wasstirred at room temperature for 3 days. As low transformation wasnoticed, additional formaldehyde (37% solution in water; 252 μL; 3.37mmol) was added and the reaction mixture was stirred at 70° C. for 16hours.

H₂O and EtOAc were added. The organic layer was decanted, dried overMgSO₄, filtered and evaporated to dryness.

The residue (0.325 g) was purified by silica gel chromatography(irregular SiOH, 40 g, mobile phase: gradient from 95% DCM, 5% MeOH,0.5% NH₄OH to 90% DCM, 10% MeOH, 1% NH₄OH). The fractions containing theproduct were mixed and concentrated to afford an intermediate fraction(106 mg) which was crystallized from a mixture of Et₂O/ACN to affordafter filtration and drying 86 mg (28%) of compound 6. MP: 170° C. (K)

EXAMPLE B6

Preparation of Compound 7

as a HCl Salt (1.65HCl 2.2H₂O)

A solution of intermediate 4 (293 mg; 0638 mmol), formaldehyde (37%solution in water; 143 μL; 1.91 mmol) in 1,4-dioxane (5.16 mL) wasstirred at room temperature for 3 days. As no transformation wasnoticed, additional formaldehyde (37% solution in water; 238 μL; 3.18mmol) was added and the reaction mixture was stirred at 70° C. for 16hours. Again, additional formaldehyde (37% solution in water; 477 μL;6.36 mmol) was added and the reaction mixture was stirred at 70° C. for16 hours. H₂O and EtOAc were added. The organic layer was decanted,dried over MgSO₄, filtered and evaporated to dryness.

The residue (0.48 g) was purified by silica gel chromatography(Spherical bare silica 5 μm 150×30.0 mm, Mobile phase: Gradient from0.2% NH₄OH, 98% DCM, 2% MeOH to 1% NH₄OH, 90% DCM, 10% MeOH). Thefractions containing the product were mixed and concentrated to afford148 mg of an intermediate fraction which was purified by achiral SFC(Stationary phase: CYANO 6 μm 150×21.2 mm, Mobile phase: 90% CO₂, 10%MeOH (0.3% iPrNH₂)). The fractions containing the product were mixed andconcentrated to afford 100 mg of an intermediate fraction which wasdissolved in MeOH. 0.1 mL of HCl in iPrOH (2-5N) was added at 0° C. Themixture was then concentrated and the resulting residue was taken upwith Et₂O. The precipitate was filtered and dried to afford 103 mg (29%)of compound 7 as a red solid. MP: 152° C. (K)

EXAMPLE B7

Preparation of Compound 8

A solution of intermediate 11 (382 mg; 0.82 mmol) and formaldehyde (37%solution in water; 308 μL; 4.11 mmol) in dioxane (10 mL) was heated at60° C. for 3 days. H₂O and EtOAc were added. The organic layer wasdecanted, dried over MgSO₄, filtered and evaporated to dryness. Theresidue was purified by chromatography over silica gel (Spherical baresilica 5 μm 150×30.0 mm; gradient: from 71% heptane, 1% MeOH (+10%NH₄OH), 28% EtOAc to 0% heptane, 20% MeOH (+10% NH₄OH), 80% EtOAc). Thepure fractions were collected and evaporated to dryness. The residue (65mg) was purified by reverse phase chromatography (X-Bridge-C18 5 μm30*150 mm; gradient: from 80% NH₄HCO₃ 0.5%, 20% CH₃CN to 0% NH₄HCO₃0.5%, 100% CH₃CN). The pure fractions were collected and evaporated togive 15 mg (4%) of compound 8. MP: 266° C. (K).

EXAMPLE B8

Preparation of Compound 9

A solution of intermediate 5 (0.21 g; 0.46 mmol) and formaldehyde (37%solution in water; 0.1 mL; 1.4 mmol) in 1,4-dioxane (8 mL) was stirredat room temperature for 3 days. After a week, additional formaldehyde(37% solution in water; 0.5 mL; 20.55 mmol) was added, the mixture wasstirred at room temperature for further 2 days. H₂O and EtOAc wereadded. The organic layer was extracted, dried over MgSO₄ and evaporatedto dryness.

The resulting residue (170 mg) was purified by reverse phase (Stationaryphase: X-Bridge-C18 5 μm 30*150 mm, Mobile phase: Gradient from 85%NH₄HCO₃ 0.5%, 15% ACN to 0% NH₄HCO₃ 0.5%, 100% ACN). The fractionscontaining the product were mixed and concentrated to afford anintermediate fraction (10 mg) which was freeze-dried withacetonitrile/water 20/80 to give 9 mg (4%) of compound 9 as a yellowpowder. MP: gum at 80° C. (K).

Analytical Part

LCMS (Liquid Chromatography/Mass Spectrometry) (See Table A1)

The LC measurement was performed using a UPLC (Ultra Performance LiquidChromatography) Acquity (Waters) system comprising a binary pump withdegasser, an autosampler, a diode-array detector (DAD) and a column asspecified in the respective methods below, the column is hold at atemperature of 40° C. Flow from the column was brought to a MS detector.The MS detector was configured with an electrospray ionization source.The capillary needle voltage was 3 kV and the source temperature wasmaintained at 130° C. on the Quattro (triple quadrupole massspectrometer from Waters). Nitrogen was used as the nebulizer gas. Dataacquisition was performed with a Waters-Micromass MassLynx-Openlynx datasystem. Reversed phase UPLC was carried out on a Waters Acquity BEH(bridged ethylsiloxane/silica hybrid) C18 column (1.7 μm, 2.1×100 mm)with a flow rate of 0.343 ml/min. Two mobile phases (mobile phase A: 95%7 mM ammonium acetate/5% acetonitrile; mobile phase B: 100%acetonitrile) were employed to run a gradient condition from 84.2% A and15.8% B (hold for 0.49 minutes) to 10.5% A and 89.5% B in 2.18 minutes,hold for 1.94 min and back to the initial conditions in 0.73 min, holdfor 0.73 minutes. An injection volume of 2 μl was used. Cone voltage was20V for positive and negative ionization mode. Mass spectra wereacquired by scanning from 100 to 1000 in 0.2 seconds using an interscandelay of 0.1 seconds.

DSC

For a number of compounds, melting points (MP) were determined with aDSC1 (Mettler-Toledo). Melting points were measured with a temperaturegradient of 10° C./minute. Maximum temperature was 350° C. Values arepeak values.

For a number of compounds, melting points were obtained with a Koflerhot bench, consisting of a heated plate with linear temperaturegradient, a sliding pointer and a temperature scale in degrees Celsius.

NMR

For compound 1, 2, 6 to 9, the NMR experiments were carried out using aBruker Avance III 500 using internal deuterium lock and equipped with aninverse triple-resonance (¹H, ¹³C, ¹⁵N TXI) probe head. Chemical shifts(δ) are reported in parts per million (ppm).

For compound 3 and 4, each fraction was dissolved in 250 μl ofwater-free DMSO-d6 and the resulting solution was transferred into a 5mm Shigemi NMR tube with a magnetic susceptibility matched for therespective solvent.

Experiments were recorded on a Bruker Avance 600 MHz spectrometerequipped with a inverse detection 5-mm cryoprobe (CPTCI). 1D ¹H and 2DNOESY, HSQC and HMBC spectra were recorded running standard Bruker pulseprograms. The NOESY spectrum was used for the determination ofthrough-space connectivities; the HMBC spectrum for through-bondconnectivities. Chemical shifts (δ) are reported in ppm. ¹H NMR chemicalshift data were obtained from the 1D ¹H spectrum using the centre of theDMSO-d5 multiplet at 2.50 ppm or the centre of the acetonitrile-d2multiplet at 1.94 ppm as internal reference. Coupling constants aremeasured in Hz. ¹³C NMR chemical shifts were obtained using the centreof the DMSO-d6 multiplet at 39.51 ppm as internal reference.

TABLE A1 Co. No. means compound number; Retention time (R_(t)) inminutes; MP means melting point (° C.). As understood by a personskilled in the art, compounds synthesized using the protocols asindicated may exist as a solvate e.g. hydrate, and/or contain residualsolvent or minor impurities. Co. (Kofler(K) No. Compound MP or DSC)R_(t) [M + H]⁺ 1

190° C. DSC 2.46 (98.1% purity) 459 2

 80° C. (gummy) K 2.29 (94.3% purity) 445 3

255° C. K 2.07 (100% purity) 445 4

256° C. K 2.06 (100% of purity) 445 5

6

170° C. K 2.47 (99% of purity) 445 7

152° C. (gummy) K 2.64 (95% of purity) 473 8

266° C. K 2.58 (95% of purity) 477 9

 80° C. (gummy) K 2.23 (100% of purity) 473

Compound 1

¹H NMR was performed at 350° K

¹H NMR (500 MHz, DMSO-d₆) δ ppm 0.99 (d, J=6.5 Hz, 6 H) 2.84 (spt, J=6.5Hz, 1 H) 2.88-2.93 (m, 2 H) 3.56 (br. s., 2 H) 3.76 (s, 3 H) 3.82-3.91(m, 5 H) 3.93 (s, 3 H) 6.42 (d, J=2.2 Hz, 1 H) 6.57 (d, J=2.2 Hz, 1 H)6.97 (d, J=2.7 Hz, 1 H) 7.26 (dd, J=9.1, 2.7 Hz, 1 H) 7.75 (d, J=9.1 Hz,1 H) 8.14 (s, 1 H) 8.46 (s, 1 H) 8.87 (s, 1 H)

Compound 2

¹H NMR was performed at 350° K

¹H NMR (500 MHz, DMSO-d₆) δ ppm 0.99 (d, J=6.6 Hz, 6 H) 2.84 (spt, J=6.6Hz, 1 H) 2.88-2.95 (m, 2 H) 3.56 (br. s., 2 H) 3.76 (s, 3 H) 3.80-3.95(m, 5 H) 6.43 (d, J=2.2 Hz, 1 H) 6.57 (d, J=2.2 Hz, 1 H) 6.99 (d, J=2.7Hz, 1 H) 7.26 (dd, J=9.5, 2.7 Hz, 1 H) 7.75 (d, J=9.5 Hz, 1 H) 8.35 (br.s., 2 H) 8.92 (s, 1 H) 13.08 (br. s., 1 H)

Compound 3

¹H NMR was performed at 300° K

¹H NMR (600 MHz, DMSO-d₆) δ ppm 0.98 (d, J=6.42 Hz, 6 H) 2.82 (spt,J=6.50 Hz, 1 H) 2.88 (t, J=4.53 Hz, 2 H) 3.69 (s, 3 H) 3.91 (s, 3 H)6.29 (d, J=2.27 Hz, 1 H) 6.42 (d, J=2.27 Hz, 1 H) 6.89 (br. s., 1 H)7.25 (br. s., 1 H) 7.75 (d, J=9.07 Hz, 1 H) 8.18 (s, 1 H) 8.53 (s, 1 H)8.89 (s, 1 H)

Compound 4

¹H NMR was performed at 300° K

¹H NMR (600 MHz, DMSO-d₆) δ ppm 0.96 (d, J=6.70 Hz, 6 H) 2.81 (spt,J=6.70 Hz, 1 H) 2.86 (t, J=4.34 Hz, 2 H) 3.79 (s, 3 H) 3.91 (s, 3 H)6.26 (d, J=1.89 Hz, 1 H) 6.41 (d, J=2.27 Hz, 1 H) 6.91 (br. s., 1 H)7.27 (br. s., 1 H) 7.75 (d, J=9.44 Hz, 1 H) 8.18 (s, 1 H) 8.53 (s, 1 H)8.89 (s, 1 H)

Compound 6

¹H NMR was performed at 300° K

¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.43 (t, J=7.3 Hz, 3 H) 2.22 (br s, 3 H)2.82 (br s, 2 H) 3.50-4.10 (m, 10 H) 4.20 (q, J=7.3 Hz, 2 H) 6.46 (d,J=1.9 Hz, 1 H) 6.59 (d, J=1.9 Hz, 1 H) 6.92 (br s, 1 H) 7.25 (br d,J=7.3 Hz, 1 H) 7.77 (d, J=9.1 Hz, 1 H) 8.20 (s, 1 H) 8.58 (s, 1 H) 8.92(s, 1 H)

Compound 7

¹H NMR was performed at 300° K

¹H NMR (500 MHz, DMSO-d₆) δ ppm 1.32 (dd, J=9.8, 6.6 Hz, 6 H) 1.44 (t,J=7.4 Hz, 3 H) 3.35-3.60 (m, 3 H) 3.93 (s, 3 H) 3.79 (s, 3 H) 4.22 (q,J=7.5 Hz, 2 H) 4.43 (br d, J=12.9 Hz, 1 H) 4.58 (br s, 1 H) 6.56 (d,J=2.5 Hz, 1 H) 6.70 (d, J=2.2 Hz, 1 H) 7.17 (br d, J=1.9 Hz, 1 H) 7.30(dd, J=9.3, 2.4 Hz, 1 H) 7.84 (d, J=9.1 Hz, 1 H) 8.23 (s, 1 H) 8.62 (s,1 H) 9.02 (s, 1 H) 10.26 (br s, 1 H)

Compound 8

¹H NMR was performed at 350° K

¹H NMR (500 MHz, DMSO-d₆) δ ppm 0.99 (d, J=6.6 Hz, 6 H) 2.85 (spt, J=6.5Hz, 1 H) 2.92 (t, J=4.6 Hz, 2 H) 3.58 (br s, 2 H) 3.80-4.05 (m, 11 H)6.84 (d, J=6.9 Hz, 1 H) 6.92 (br s, 1 H) 7.20 (br d, J=9.5 Hz, 1 H) 7.79(d, J=9.1 Hz, 1 H) 8.15 (s, 1 H) 8.46 (s, 1 H) 8.89 (s, 1 H)

Compound 9

¹H NMR was performed at 300° K

¹H NMR (500 MHz, DMSO-d₆) δ ppm 0.97 (br d, J=5.4 Hz, 6 H) 1.64 (br s, 2H) 2.72 (br s, 2 H) 2.84 (spt, J=6.4 Hz, 1 H) 3.50-3.80 (m, 7 H) 3.84(s, 3 H) 3.92 (s, 3 H) 6.21 (d, J=2.2 Hz, 1 H) 6.61 (d, J=2.5 Hz, 1 H)6.92 (br s, 2 H) 7.71 (d, J=9.5 Hz, 1 H) 8.19 (s, 1 H) 8.54 (s, 1 H)8.91 (s, 1 H)

Some signals of the diazepine ring are broadened beyond detection inspectra measured at 300 K in DMSO-d6.

Pharmacological Part

Biological Assays a

FGFR1 (Enzymatic Assay)

In a final reaction volume of 30 μL, FGFR1 (h) (25 ng/ml) was incubatedwith 50 mM HEPES pH 7.5, 6 mM MnCl₂, 1 mM DTT, 0.1 mM Na₃VO₄, 0.01%Triton-X-100, 500 nM Btn-Flt3 and 5 μM ATP in the presence of compound(1% DMSO final). After incubation for 60 minutes at room temperature thereaction was stopped with 2.27 nM EU-anti P-Tyr, 7 mM EDTA, 31.25 nMSA-XL-665 and 0.02% BSA which was present for 60 minutes at roomtemperature. Time-Resolved Fluorescence Resonance Energy Transfer(TR-FRET) signal (ex340 nm. Em 620 nm, em 655 nm) was measuredafterwards and results are expressed in RFU (Relative FluorescenceUnits). In this assay, the inhibitory effect of different compoundconcentrations (range 10 μM to 0.1 nM) was determined and used tocalculate an IC₅₀ (M) and pIC₅₀ (−logIC₅₀) value.

FGFR2 (Enzymatic Assay)

In a final reaction volume of 30 μL, FGFR2 (h) (150 ng/ml) was incubatedwith 50 mM HEPES pH 7.5, 6 mM MnCl₂, 1 mM DTT, 0.1 mM Na₃VO₄, 0.01%Triton-X-100, 500 nM Btn-Flt3 and 0.4 μM ATP in the presence of compound(1% DMSO final). After incubation for 60 minutes at room temperature thereaction was stopped with 2.27 nM EU-anti P-Tyr, 7 mM EDTA, 31.25 nMSA-XL-665 and 0.02% BSA which was present for 60 minutes at roomtemperature. Time-Resolved Fluorescence Resonance Energy Transfer(TR-FRET) signal (ex340 nm. Em 620 nm, em 655 nm) was measuredafterwards and results are expressed in (Relative Fluorescence Units).In this assay, the inhibitory effect of different compoundconcentrations (range 10 μM to 0.1 nM) was determined and used tocalculate an IC₅₀ (M) and pIC₅₀ (−logIC₅₀) value.

FGFR3 (Enzymatic Assay)

In a final reaction volume of 30 μL, FGFR3 (h) (40 ng/ml) was incubatedwith 50 mM HEPES pH 7.5, 6 mM MnCl₂, 1 mM DTT, 0.1 mM Na₃VO₄, 0.01%Triton-X-100, 500 nM Btn-Flt3 and 25 μM ATP in the presence of compound(1% DMSO final). After incubation for 60 minutes at room temperature thereaction was stopped with 2.27 nM EU-anti P-Tyr, 7 mM EDTA, 31.25 nMSA-XL-665 and 0.02% BSA which was present for 60 minutes at roomtemperature. Time-Resolved Fluorescence Resonance Energy Transfer(TR-FRET) signal (ex340 nm. Em 620 nm, em 655 nm) was measuredafterwards and results are expressed in RFU (Relative FluorescenceUnits). In this assay, the inhibitory effect of different compoundconcentrations (range 10 μM to 0.1 nM) was determined and used tocalculate an IC₅₀ (M) and pIC₅₀ (−logIC₅₀) value.

FGFR4 (Enzymatic Assay)

In a final reaction volume of 30 μL, FGFR4 (h) (60 ng/ml) was incubatedwith 50 mM HEPES pH 7.5, 6 mM MnCl₂, 1 mM DTT, 0.1 mM Na₃VO₄, 0.01%Triton-X-100, 500 nM Btn-Flt3 and 5 μM ATP in the presence of compound(1% DMSO final). After incubation for 60 minutes at room temperature thereaction was stopped with 2.27 nM EU-anti P-Tyr, 7 mM EDTA, 31.25 nMSA-XL-665 and 0.02% BSA which was present for 60 minutes at roomtemperature. Time-Resolved Fluorescence Resonance Energy Transfer(TR-FRET) signal (ex340 nm. Em 620 nm, em 655 nm) was measuredafterwards and results are expressed in RFU (Relative FluorescenceUnits). In this assay, the inhibitory effect of different compoundconcentrations (range 10 μM to 0.1 nM) was determined and used tocalculate an IC₅₀ (M) and pIC₅₀ (−logIC₅₀) value.

KDR (VEGFR2) (Enzymatic Assayl

In a final reaction volume of 30 μL, KDR (h) (150 ng/ml) was incubatedwith 50 mM HEPES pH 7.5, 6 mM MnCl₂, 1 mM DTT, 0.1 mM Na₃VO₄, 0.01%Triton-X-100, 500 nM Btn-Flt3 and 3 μM ATP in the presence of compound(1% DMSO final). After incubation for 120 minutes at room temperaturethe reaction was stopped with 2.27 nM EU-anti P-Tyr, 7 mM EDTA, 31.25 nMSA-XL-665 and 0.02% BSA which was present for 60 minutes at roomtemperature. Time-Resolved Fluorescence Resonance Energy Transfer(TR-FRET) signal (ex340 nm. Em 620 nm, em 655 nm) was measuredafterwards and results are expressed in RFU (Relative FluorescenceUnits). In this assay, the inhibitory effect of different compoundconcentrations (range 10 μM to 0.1 nM) was determined and used tocalculate an IC₅₀ (M) and pIC₅₀ (−logIC₅₀) value.

Ba/F3-FGFR1 (Minus IL3 or Plus IL3) (Cellular Proliferation Assay)

In a 384 well plate, 100 nl of compound dilution in DMSO was sprayedbefore adding 50 μl cell culture medium (phenol red free RPMI-1640, 10%FBS, 2 mM L-Glutamine and 50 μg/ml Gentamycin) containing 20000 cellsper well of Ba/F3-FGFR1-transfected cells. Cells were put in anincubator at 37° C. and 5% CO₂. After 24 hours, 10 μl of Alamar Bluesolution (0.5 mM K₃Fe(CN)₆, 0.5 mM K₄Fe(CN)₆, 0.15 mM Resazurin and 100mM Phosphate Buffer) was added to the wells, incubated for 4 hours at37° C. and 5% CO₂ before RFU's (Relative Fluorescence Units) (ex. 540nm., em. 590 nm.) were measured in a flurorescence plate reader.

In this assay, the inhibitory effect of different compoundconcentrations (range 10 μM to 0.1 nM) was determined and used tocalculate an IC₅₀ (M) and pIC₅₀ (−logIC₅₀) value. As a counterscreen thesame experiment was performed in the presence of 10 ng/ml murine IL3.

Ba/F3-FGFR3 (Minus IL3 or Plus IL3) (Cellular Proliferation Assay)

In a 384 well plate, 100 nl of compound dilution in DMSO was sprayedbefore adding 50 μl cell culture medium (phenol red free RPMI-1640, 10%FBS, 2 mM L-Glutamine and 50 μg/ml Gentamycin) containing 20000 cellsper well of Ba/F3-FGFR3-transfected cells. Cells were put in anincubator at 37° C. and 5% CO₂. After 24 hours, 10 μl of Alamar Bluesolution (0.5 mM K₃Fe(CN)₆, 0.5 mM K₄Fe(CN)₆, 0.15 mM Resazurin and 100mM Phosphate Buffer) was added to the wells, incubated for 4 hours at37° C. and 5% CO₂ before RFU's (Relative Fluorescence Units) (ex. 540nm., em. 590 nm.) were measured in a flurorescence plate reader.

In this assay, the inhibitory effect of different compoundconcentrations (range 10 μM to 0.1 nM) was determined and used tocalculate an IC₅₀ (M) and pIC₅₀ (−logIC₅₀) value.

As a counterscreen the same experiment was performed in the presence of10 ng/ml murine IL3.

Ba/F3-KDR (Minus IL3 or Plus IL3) (Cellular Proliferation Assay)

In a 384 well plate, 100 nl of compound dilution in DMSO was sprayedbefore adding 50 μl cell culture medium (phenol red free RPMI-1640, 10%FBS, 2 mM L-Glutamine and 50 μg/ml Gentamycin) containing 20000 cellsper well of Ba/F3-KDR-transfected cells. Cells were put in an incubatorat 37° C. and 5% CO₂. After 24 hours, 10 μl of Alamar Blue solution (0.5mM K₃Fe(CN)₆, 0.5 mM K₄Fe(CN)₆, 0.15 mM Resazurin and 100 mM PhosphateBuffer) was added to the wells, incubated for 4 hours at 37° C. and 5%CO₂ before RFU's (Relative Fluorescence Units) (ex. 540 nm., em. 590nm.) were measured in a flurorescence plate reader.

In this assay, the inhibitory effect of different compoundconcentrations (range 10 μM to 0.1 nM) was determined and used tocalculate an IC₅₀ (M) and pIC₅₀ (−logIC₅₀) value.

As a counterscreen the same experiment was performed in the presence of10 ng/ml murine IL3.

Ba/F3-FGFR4 (Cellular Proliferation Assay)

In a 384 well plate, 100 nl of compound dilution in DMSO was sprayedbefore adding 50 μl cell culture medium (phenol red free RPMI-1640, 10%FBS, 2 mM L-Glutamine and 50 μg/ml Gentamycin) containing 20000 cellsper well of Ba/F3-FGFR4-transfected cells. Cells were put in anincubator at 37° C. and 5% CO₂. After 24 hours, 10 μl of Alamar Bluesolution (0.5 mM K₃Fe(CN)₆, 0.5 mM K₄Fe(CN)₆, 0.15 mM Resazurin and 100mM Phosphate Buffer) was added to the wells, incubated for 4 hours at37° C. and 5% CO₂ before RFU's (Relative Fluorescence Units) (ex. 540nm., em. 590 nm.) were measured in a flurorescence plate reader.

In this assay, the inhibitory effect of different compoundconcentrations (range 10 μM to 0.1 nM) was determined and used tocalculate an IC₅₀ (M) and pIC₅₀ (−logIC₅₀) value.

Data for the compounds of the invention in the above assays are providedin Table A2.

TABLE A2 BAF3- BAF3- BAF3- BAF3- BAF3- BAF3- FGFR1 FGFR1 FGFR3 FGFR3 KDRKDR VEGFR (MIN (PLUS (MIN (PLUS (MIN (PLUS BAF3- Comp. FGFR1 FGFR2 FGFR3FGFR4 KDR IL3 IL3 IL3 IL3 IL3 IL3 FGFR4 No. pIC50 pIC50 pIC50 pIC50pIC50 pIC50) pIC50) pIC50) pIC50) pIC50) pIC50) (pIC50) 1 6.3 6.5 6.15.3 5.6 5.2 <5 ~5.0 <5 <5 <5 5.0Biological Assays BEnzyme Binding Assays (KINOMEscan®)

Kinase enzyme binding affinities of compounds disclosed herein weredetermined using the KINOMEscan® technology performed by DiscoveRxCorporation, San Diego, Calif., USA (www.kinomescan.com). Table A3reports the obtained Kd values (nM), with the Kd being the inhibitorbinding constant:

TABLE A3 Kd Kd Kd Kd Kd FGFR1 FGFR2 FGFR3 FGFR4 VEGFR2 Compound (nM)(nM) (nM) (nM) (nM) 1 314 674 325 778 >3010 2 17 63 68 68 741 3 720 620300 1900 >3000 4 740 900 870 >3000 >3000 6 2400 2900 2200 >3000 >3000 779 340 170 230 1700 9 54 117 138 355 922

The invention claimed is:
 1. A method of inhibiting a FGFR kinase, whichmethod comprises contacting the kinase with a kinase-inhibiting compoundselected from the group consisting of a compound of formula (I):

a tautomeric form, and stereochemically isomeric form thereof, wherein nrepresents an integer equal to 1 or 2; R₁ represents hydrogen,C₁₋₆alkyl, hydroxyC₁₋₆alkyl, C₁₋₆alkyl substituted with —C(═O)NHCH₃, orC₁₋₆alkyl substituted with —S(═O)₂- C₁₋₄alkyl; R_(2a) representshydrogen, fluoro or chloro; R_(2b) or R_(2c) each independentlyrepresent methoxy or hydroxyl; R₃ represent hydrogen, C₁₋₆alkyl,C₃₋₆cycloalkyl, or C₁₋₂alkyl substituted with C₃₋₆cycloalkyl; R₄represents hydrogen, methyl or ethyl; or a pharmaceutically acceptablesalt thereof or a solvate thereof.
 2. A method according to claim 1,wherein the compound is


3. A method according to claim 1, wherein the compound has the followingstructure

or a pharmaceutically acceptable salt thereof or a solvate thereof.
 4. Amethod according to claim 1, wherein R_(2a) represents hydrogen orfluoro.
 5. A method according to claim 4, wherein R_(2a) representsfluoro.
 6. A method according to claim 1, wherein the compound has thefollowing structure

or a pharmaceutically acceptable salt thereof or a solvate thereof.
 7. Amethod according to claim 1, wherein n represents an integer equal to 1.8. A method according to claim 1, wherein R₃ represents hydrogen.
 9. Amethod according to claim 1, wherein R₃ represents C₁₋₆alkyl.
 10. Amethod according to claim 1, wherein the compound has the followingstructure

or a pharmaceutically acceptable salt thereof or a solvate thereof. 11.A method according to claim 1, wherein R₁ represents hydrogen orC₁₋₆alkyl.
 12. A method according to claim 11, wherein R₁ representsC₁₋₆alkyl.
 13. A method according to claim 12, wherein R₁ representsmethyl.
 14. A method according to claim 1, wherein R_(2b) representsmethoxy.
 15. A method according to claim 1, wherein R_(2b) representshydroxy.
 16. A method according to claim 1, wherein R_(2c) representsmethoxy.
 17. A method according to claim 1, wherein R_(2c) representshydroxy.
 18. A method according to claim 1, wherein the compound is

or a pharmaceutically acceptable salt or solvate thereof.
 19. A methodaccording to claim 1, wherein the compound is selected from

or a pharmaceutically acceptable salt thereof or a solvate thereof.